Short protocols in protein science: a compendium of methods from "Current protocols in protein science"
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
[Hoboken. N.J.]
Wiley
2003
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| Series: | Current protocols
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| Subjects: | |
| Online Access: | Publisher description Table of contents Inhaltsverzeichnis |
| Item Description: | Includes bibliographical references and index |
| Physical Description: | Getr. Zählung Ill., graph. Darst. |
| ISBN: | 0471483389 |
Staff View
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| 245 | 1 | 0 | |a Short protocols in protein science |b a compendium of methods from "Current protocols in protein science" |c ed. by John E. Coligan ... |
| 264 | 1 | |a [Hoboken. N.J.] |b Wiley |c 2003 | |
| 300 | |a Getr. Zählung |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
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| 338 | |b nc |2 rdacarrier | ||
| 490 | 0 | |a Current protocols | |
| 500 | |a Includes bibliographical references and index | ||
| 650 | 7 | |a Eiwitsynthese |2 gtt | |
| 650 | 7 | |a Eiwitten |2 gtt | |
| 650 | 7 | |a Proteínas (análise;isolamento e purificação) |2 larpcal | |
| 650 | 4 | |a Protéines - Purification - Manuels de laboratoire | |
| 650 | 7 | |a Scheidingstechnieken |2 gtt | |
| 650 | 7 | |a Spectrometrie |2 gtt | |
| 650 | 4 | |a Proteins |x Purification |v Laboratory manuals | |
| 650 | 4 | |a Proteins |x analysis |v Laboratory Manuals | |
| 650 | 4 | |a Proteins |x isolation & purification |v Laboratory Manuals | |
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| 856 | 4 | |u http://www.loc.gov/catdir/toc/wiley031/2003012325.html |3 Table of contents | |
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Record in the Search Index
| _version_ | 1804132785717248000 |
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| adam_text | SHORT PROTOCOLS IN PROTEIN SCIENCE A COMPENDIUM OF METHODS FROM CURRENT
PROTOCOLS IN PROTEIN SCIENCE EDITORIAL BOARD JOHN E. COLIGAN NATIONAL
INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES, ROCKVILLE, MARYLAND BEN M.
DUNN UNIVERSITY OF FLORIDA, GAINESVILLE, FLORIDA DAVID W. SPEICHER THE
WISTAR INSTITUTE, PHILADELPHIA, PENNSYLVANIA PAUL T. WINGFIELD NATIONAL
INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES, BETHESDA,
MARYLAND UNIVERSITATS- :.IND LA-NDESBIBLIOTHCL; BIBLIOTHEK BIOLOGIC
SCHNITTSPAHNSTR. 10, 64287 DARMSTADT *******... ,4.,.. »* WILEY
PUBLISHED BY JOHN WILEY & SONS, INC. 52509556 CONTENTS PREFACE XVII
CONTRIBUTORS XXI 1 STRATEGIES OF PROTEIN PURIFICATION AND
CHARACTERIZATION 1-1 1.1 OVERVIEW OF PROTEIN PURIFICATION AND
CHARACTERIZATION 1-1 AIMS AND OBJECTIVES 1-1 SOURCES OF MATERIAL FOR
PROTEIN PURIFICATION 1-2 DETECTION AND ASSAY OF PROTEINS 1-3 METHODS FOR
SEPARATION AND PURIFICATION OF PROTEINS 1-3 CHARACTERIZATION OF THE
PROTEIN PRODUCT 1-5 THE PROTEIN PURIFICATION LABORATORY 1-5 1.2 PROTEIN
PURIFICATION FLOW CHARTS 1-6 SOLUBLE RECOMBINANT PROTEINS 1-6 INSOLUBLE
RECOMBINANT PROTEINS 1-8 SOLUBLE NONRECOMBINANT PROTEINS 1-9
MEMBRANE-ASSOCIATED AND INSOLUBLE NONRECOMBINANT PROTEINS 1-12 2
COMPUTATIONAL ANALYSIS 2-1 2.1 HYDROPHOBICITY PROFILES FOR PROTEIN
SEQUENCE ANALYSIS 2-1 METHODOLOGY 2-2 APPLICATIONS 2-4 CONCLUSIONS 2-6
2.2 PROTEIN SECONDARY STRUCTURE PREDICTION 2-6 METHODS FOR SECONDARY
STRUCTURE PREDICTION 2-6 APPLICATION OF SECONDARY STRUCTURE PREDICTION
METHODS 2-9 2.3 SEQUENCE SIMILARITY SEARCHING USING THE BLAST FAMILY OF
PROGRAMS 2-11 ACCESSING BLAST PROGRAMS AND DOCUMENTATION 2-11
INTRODUCTION TO BLAST 2-12 BLAST PROGRAMS 2-14 2.4 PROTEIN DATABASES ON
THE INTERNET 2-16 PROTEIN STRUCTURE DATABASES 2-16 PROTEIN FAMILY
DATABASES 2-19 2.5 PROTEIN TERTIARY STRUCTURE PREDICTION 2-20 HOMOLOGY
MODELING , 2-20 SEQUENCE PROFILE METHODS 2-22 THREADING 2-22 AB INITIO
PREDICTION 2-23 2.6 PROTEIN TERTIARY STRUCTURE MODELING 2-23 GLOSSARY
2-23 ACCESSING SWISSMODEL.PROGRAMS AND DOCUMENTATION 2-24 EXPDB DATABASE
2-24 FORMATTING A FIRST APPROACH MODELING REQUEST 2-25 VIEWING
SWISSMODEL RESULTS 2-25 2.7 COMPARATIVE PROTEIN STRUCTURE PREDICTION
2-26 STEPS IN COMPARATIVE MODELING 2-26 CONTENTS SHORT PROTOCOLS IN
PROTEIN SCIENCE 3 DETECTION AND ASSAY METHODS 3-1 3.1 SPECTROPHOTOMETRIC
DETERMINATION OF PROTEIN CONCENTRATION 3-2 BASIC PROTOCOL 1: CALCULATION
OF THE MOLAR ABSORPTION COEFFICIENT OF A PROTEIN 3-2 BASIC PROTOCOL 2:
DETERMINATION OF THE MOLAR ABSORPTION COEFFICIENT FOR A FOLDED PROTEIN
3-3 BASIC PROTOCOL 3: DETERMINATION OF PROTEIN CONCENTRATION BY
ABSORBANCE SPECTROSCOPY USING THE MOLAR ABSORPTION COEFFICIENT 3-4 BASIC
PROTOCOL 4: DETERMINATION OF PROTEIN CONCENTRATION BY ABSORPTION
SPECTROSCOPY AT 205 NM 3-4 BASIC PROTOCOL 5: DETERMINATION OF TOTAL
PROTEIN CONCENTRATION IN CRUDE PROTEIN EXTRACTS 3-5 3.2 QUANTITATIVE
AMINO ACID ANALYSIS 3-6 SAMPLE PREPARATION 3-6 CALCULATION OF THE
AVERAGE COMPOSITION 3-6 3.3 IN VITRO RADIOLABELING OF PEPTIDES AND
PROTEINS 3-8 BASIC PROTOCOL 1: IODINATION AT TYROSINE OR HISTIDINE
RESIDUES USING IODO-BEADS 3-8 ALTERNATE PROTOCOL 1: IODINATION AT
TYROSINE OR HISTIDINE RESIDUES USING CHLORAMINE T OR IODOGEN 3-10
ALTERNATE PROTOCOL 2: IODINATION AT TYROSINE OR HISTIDINE RESIDUES USING
LACTOPEROXIDASE 3-11 SUPPORT PROTOCOL 1: EQUIMOLAR IODINATION TO GIVE
HIGH YIELDS OF PRODUCT 3-11 SUPPORT PROTOCOL 2: SEPARATION OF UNLABELED
PEPTIDE FROM IODINATED PRODUCTS BY HPLC 3-12 BASIC PROTOCOL 2:
IODINATION AT LYSINE RESIDUES OR N-TERMINUS USING BOLTON-HUNTER REAGENT
3-13 BASIC PROTOCOL 3: I4 C OR 3 H LABELING AT LYSINE RESIDUES OR
N-TERMINUS BY ACETYLATION USING ANHYDRIDE 3-14 ALTERNATE PROTOCOL 3: 4
C OR 3 H LABELING AT LYSINE RESIDUES OR N-TERMINUS BY REDUCTIVE
ALKYLATION 3-15 ALTERNATE PROTOCOL 4: C OR 3 H LABELING AT CYSTEINE
RESIDUES USING IODOACETIC ACID OR IODOACETAMIDE C 3-15 BASIC PROTOCOL 4:
INTRODUCTION OF LABELED AMINO ACID RESIDUES DURING PEPTIDE SYNTHESIS
3-16 ALTERNATE PROTOCOL 5: SELECTIVE LABELING ON N-TERMINUS DURING
PEPTIDE SYNTHESIS 3-17 3.4 ASSAYS FOR TOTAL PROTEIN 3-18 BASIC PROTOCOL
1: BIURET ASSAY FOR QUANTITATION OF TOTAL PROTEIN 3-18 BASIC PROTOCOL 2:
HARTREE-LOWRY ASSAY FOR QUANTITATION OF TOTAL PROTEIN 3-20 BASIC
PROTOCOL 3: BICINCHONINIC ACID (BCA) ASSAY FOR QUANTITATION OF TOTAL
PROTEIN 3-21 BASIC PROTOCOL 4: ACID DIGESTION-NINHYDRIN METHOD FOR
QUANTITATION OF TOTAL PROTEIN 3-21 SUPPORT PROTOCOL 1: HEAT SEALING
GLASS TUBES 3-22 BASIC PROTOCOL 5: COOMASSIE DYE-BINDING ASSAY (BRADFORD
ASSAY) TO MEASURE TOTAL PROTEIN 3-23 SUPPORT PROTOCOL 2: GEL DIALYSIS OF
PROTEIN SAMPLES 3-23 SUPPORT PROTOCOL 3: TRICHLOROACETIC ACID
PRECIPITATION OF PROTEIN SAMPLES 3-24 3.5 BIOTINYLATION OF PROTEINS IN
SOLUTION AND ON CELL SURFACES 3-25 BASIC PROTOCOL 1: COVALENT ATTACHMENT
OF BIOTIN TO LYSINES 3-25 BASIC PROTOCOL 2: COVALENT ATTACHMENT OF
BIOTIN TO THIOLS 3-27 SUPPORT PROTOCOL 1: REDUCTION OF DISULFIDE BONDS
3-27 SUPPORT PROTOCOL 2: DETECTION OF BIOTINYLATED PROTEINS 3-28 3.6
METABOLIC LABELING WITH AMINO ACIDS 3-29 BASIC PROTOCOL: PULSE-LABELING
OF CELLS IN SUSPENSION WITH [ 35 S]METHIONINE 3-29 ALTERNATE PROTOCOL 1:
PULSE-LABELING OF ADHERENT CELLS WITH [ 35 S]METHIONINE 3-30 ALTERNATE
PROTOCOL 2: PULSE-CHASE LABELING OF CELLS WITH [ 33 S]METHIONINE 3-30
ALTERNATE PROTOCOL 3: LONG-TERM LABELING OF CELLS WITH [ 35 S]METHIONINE
3-31 SUPPORT PROTOCOL: TCA PRECIPITATION TO DETERMINE LABEL
INCORPORATION 3-31 SHORT PROTOCOLS IN PROTEIN SCIENCE CONTENTS IV 4
EXTRACTION, STABILIZATION, AND CONCENTRATION 4-1 4.1 DESALTING,
CONCENTRATION, AND BUFFER EXCHANGE BY DIALYSIS AND ULTRAFILTRATION 4-2
BASIC PROTOCOL 1: DESALTING AND BUFFER EXCHANGE BY DIALYSIS THROUGH
REGENERATED CELLULOSE TUBING 4-2 BASIC PROTOCOL 2: CONCENTRATION OR
DIALYSIS BY ULTRAFILTRATION THROUGH ASYMMETRIC MEMBRANE DISKS 4-3
ALTERNATE PROTOCOL 1: DIAFILTRATION OR CONCENTRATION BY TANGENTIAL-FLOW
ULTRAFILTRATION 4-5 ALTERNATE PROTOCOL 2: MICROSCALE CONCENTRATION AND
DESALTING WITH CENTRIFUGAL ULTRAFILTERS 4-7 4.2 SELECTIVE
PRECIPITATION OF PROTEINS 4-8 STRATEGIC PLANNING 4-8 BASIC PROTOCOL 1:
SELECTIVE PRECIPITATION BY SALTING OUT 4-9 ALTERNATE PROTOCOL 1:
SELECTIVE PRECIPITATION BY STEPWISE SALTING OUT 4-11 BASIC PROTOCOL 2:
SELECTIVE PRECIPITATION BY ISOIONIC PRECIPITATION: COLUMN METHOD 4-12
ALTERNATE PROTOCOL 2: SELECTIVE PRECIPITATION BY ISOIONIC PRECIPITATION:
DIALYSIS METHOD 4-13 BASIC PROTOCOL 3: SELECTIVE PRECIPITATION USING C 4
AND C 5 ORGANIC COSOLVENTS 4-14 BASIC PROTOCOL 4: SELECTIVE
PRECIPITATION USING PROTEIN EXCLUSION AND CROWDING AGENTS ANDOSMOLYTES
4-14 BASIC PROTOCOL 5: SELECTIVE PRECIPITATION USING SYNTHETIC AND
SEMISYNTHETIC POLYELECTROLYTES 4-15 BASIC PROTOCOL 6: SELECTIVE
PRECIPITATION USING METALLIC AND POLYPHENOLIC HETEROPOLYANIONS 4-17 4.3
LONG-TERM STORAGE OF PROTEINS 4-17 PROTEIN AGGREGATION 4-18 CHEMICAL
DEGRADATION 4-18 STORAGE IN NONFROZEN AQUEOUS SOLUTIONS 4-19 STORAGE AS
SALTED-OUT PRECIPITATES 4-20 STORAGE AS FROZEN SOLUTIONS 4-20 STORAGE AS
FREEZE-DRIED SOLIDS 4-21 SELECTING AN APPROPRIATE STORAGE METHOD 4-23
INHIBITION OF PROTEASES 4-23 5 PRODUCTION OF RECOMBINANT PROTEINS 5-1
5.1 PRODUCTION OF RECOMBINANT PROTEINS IN ESCHERICHIA COLI 5-2 5.2
SELECTION OF ESCHERICHIA COLI EXPRESSION SYSTEMS 5-4 BASIC PROTOCOL 1:
EXPRESSION UNDER CONTROL OF P L PROMOTER: TEMPERATURE INDUCTION 5-5
BASIC PROTOCOL 2: EXPRESSION UNDER CONTROL OF TRP PROMOTER: CHEMICAL
INDUCTION 5-6 ALTERNATE PROTOCOL 1: EXPRESSION UNDER CONTROL OF LAC/TAC
PROMOTERS 5-7 ALTERNATE PROTOCOL 2: T7 RNA POLYMERASE/PROMOTER
EXPRESSION SYSTEM 5-7 SUPPORT PROTOCOL 1: STRAIN STORAGE 5-7 SUPPORT
PROTOCOL 2: PREPARATION OF SAMPLES FOR ANALYSIS BY SDS-PAGE 5-8 SUPPORT
PROTOCOL 3: SOLUBILITY ANALYSIS 5-8 SUPPORT PROTOCOL 4: PREPARATION OF
PERIPLASMIC EXTRACTS 5-9 SUPPORT PROTOCOL 5: PREPARATION OF
EXTRACELLULAR MEDIUM SAMPLES 5-10 5.3 FERMENTATION AND GROWTH OF
ESCHERICHIA COLI FOR OPTIMAL PROTEIN PRODUCTION 5-10 BASIC PROTOCOL:
PRODUCTION OF RECOMBINANT PROTEINS IN BATCH FERMENTATIONS 5-11 ALTERNATE
PROTOCOL 1: IN VIVO LABELING OF RECOMBINANT PROTEINS WITH HEAVY METAL
DERIVATIVES 5-13 ALTERNATE PROTOCOL 2: STABLE ISOTOPIC LABELING OF
RECOMBINANT PROTEINS 5-14 SUPPORT PROTOCOL 1: MONITORING GROWTH 5-15
SUPPORT PROTOCOL 2: CHECKING STERILITY 5-15 5.4 PROTEIN EXPRESSION IN
THE BACULOVIRUS SYSTEM 5-16 BASIC PROTOCOL 1: LARGE-SCALE PRODUCTION OF
VIRAL STOCK 5-16 BASIC PROTOCOL 2: DETERMINATION OF EXPRESSION KINETICS
5-17 BASIC PROTOCOL 3: PRODUCTION IN BIOREACTORS 5-18 ALTERNATE
PROTOCOL: PRODUCTION IN PERFUSION CULTURES 5-19 SUPPORT PROTOCOL:
HARVESTING 5-20 CONTENTS SHORT PROTOCOLS IN PROTEIN SCIENCE 5.5 CULTURE
OF YEAST FOR THE PRODUCTION OF HETEROLOGOUS PROTEINS 5-21 BASIC PROTOCOL
1: SMALL-SCALE EXPRESSION USING S. CEREVISIAE GALACTOSE-REGULATED
VECTORS 5-21 ALTERNATE PROTOCOL 1: EXPRESSION USING GLUCOSE-REPRESSIBLE
ADH2 VECTORS 5-23 ALTERNATE PROTOCOL 2: EXPRESSION USING VECTORS WITH
GLYCOLYTIC GENE PROMOTERS 5-23 BASIC PROTOCOL 2: SMALL-SCALE EXPRESSION
IN PICHIA PASTORIS 5-24 SUPPORT PROTOCOL: SMALL-SCALE PREPARATION OF
PROTEIN EXTRACTS 5-25 5.6 OVERVIEW OF PROTEIN EXPRESSION BY MAMMALIAN
CELLS 5-26 5.7 PRODUCTION OF RECOMBINANT PROTEINS IN MAMMALIAN CELLS
5-29 BASIC PROTOCOL 1: PLASMID PURIFICATION BY
ALKALINE-LYSIS/ANION-EXCHANGE CAPTURE 5-30 BASIC PROTOCOL 2:
TRANSFECTION OF CELLS BY LIPOFECTION 5-32 BASIC PROTOCOL 3: SELECTION
FOR NEOMYCIN PHOSPHOTRANSFERASE (NPTII) WITH GENETICIN (G418) 5-32
SUPPORT PROTOCOL: DETERMINATION OF BACKGROUND SENSITIVITY TO G418 5-33
BASIC PROTOCOL 4: AMPLIFICATION OF DIHYDROFOLATE REDUCTASE (DHFR) WITH
METHOTREXATE (MTX) ^ 5-34 BASIC PROTOCOL 5: ADAPTATION OF SUSPENSION
CELLS TO PRODUCTION MEDIUM THROUGH MULTIPLE PASSAGING 5-35 BASIC
PROTOCOL 6: GROWTH OF CELLS IN BATCH MODE IN LARGE-SCALE SPINNER CULTURE
VESSELS 5-36 BASIC PROTOCOL 7: GROWTH OF CELLS IN LARGE-SCALE BATCH
REACTORS 5-38 ALTERNATE PROTOCOL: GROWTH OF CELLS IN CONTINUOUS CULTURE
IN BIOREACTORS 5-40 BASIC PROTOCOL 8: HARVESTING A SECRETED PRODUCT FROM
SPINNER CULTURES AND LARGE-SCALE REACTORS 5-41 BASIC PROTOCOL 9:
HARVESTING A CELL-ASSOCIATED PRODUCT FROM SPINNER CULTURES AND
LARGE-SCALE REACTORS 5-43 5.8 PREPARATION OF CELL CULTURES AND VACCINIA
VIRUS STOCKS 5-44 BASIC PROTOCOL 1: CULTURE OF MONOLAYER CELLS 5-44
BASIC PROTOCOL 2: CULTURE OF CELLS IN SUSPENSION 5-45 BASIC PROTOCOL 3:
PREPARATION OF A VACCINIA VIRUS STOCK 5-46 5.9 GENERATION OF RECOMBINANT
VACCINIA VIRUSES 5-47 BASIC PROTOCOL 1: TRANSFECTION OF INFECTED CELLS
WITH A VACCINIA VECTOR 5-47 BASIC PROTOCOL 2: SELECTION AND SCREENING OF
RECOMBINANT VIRUS PLAQUES 5-52 BASIC PROTOCOL 3: AMPLIFICATION OF A
PLAQUE 5-54 5.10 CHOICE OF CELLULAR PROTEIN EXPRESSION SYSTEM 5-55 5.11
USE OF THE GATEWAY SYSTEM FOR PROTEIN EXPRESSION IN MULTIPLE HOSTS 5-61
6 PURIFICATION OF RECOMBINANT PROTEINS 6-1 6.1 OVERVIEW OF THE
PURIFICATION OF RECOMBINANT PROTEINS PRODUCED IN ESCHERICHIA COLI 6-3
DETERMINING SOLUBILITY ^ 6-3 LOCALIZING PROTEIN 6-7 ISOLATING SOLUBLE
PROTEINS 6-9 ISOLATING INSOLUBLE PROTEINS 6-10 6.2 PREPARATION OF
SOLUBLE PROTEINS FROM ESCHERICHIA COLI 6-10 BASIC PROTOCOL: PURIFICATION
OF A PROTEIN EXPRESSED IN ESCHERICHIA COLI IN A SOLUBLE STATE:
INTERLEUKIN LP 6-10 6.3 PREPARATION AND EXTRACTION OF INSOLUBLE
(INCLUSION-BODY) PROTEINS FROM ESCHERICHIA COLI 6-16 BASIC PROTOCOL 1:
PREPARATION AND EXTRACTION OF INSOLUBLE (INCLUSION-BODY) PROTEINS FROM
ESCHERICHIA COLI 6-16 BASIC PROTOCOL 2: MEDIUM-PRESSURE GEL-FILTRATION
CHROMATOGRAPHY IN THE PRESENCE OF GUANIDINE HYDROCHLORIDE 6-17 6.4
OVERVIEW OF PROTEIN FOLDING V 6-19 HOW PROTEINS FOLD 6-19 HOW TO FOLD
PROTEINS 6-21 SHORT PROTOCOLS IN PROTEIN SCIENCE CONTENTS VI 6.5 FOLDING
AND PURIFICATION OF INSOLUBLE (INCLUSION BODY) PROTEINS FROM ESCHERICHIA
COLI 6-26 BASIC PROTOCOL 1: FOLDING AND PURIFICATION OF BOVINE GROWTH
HORMONE 6-26 BASIC PROTOCOL 2: FOLDING AND PURIFICATION OF HUMAN
INTERLEUKIN 2 6-28 BASIC PROTOCOL 3: FOLDING AND PURIFICATION OF A
HISTIDINE-TAGGED PROTEIN: HIV-1 INTEGRASE 6-30 6.6 EXPRESSION AND
PURIFICATION OF GST FUSION PROTEINS 6-33 BASIC PROTOCOL 1: EXPRESSION OF
GLUTATHIONE-S-TRANSFERASE FUSION PROTEIN 6-33 BASIC PROTOCOL 2: AFFINITY
CHROMATOGRAPHY PURIFICATION OF A SOLUBLE GST FUSION PROTEIN 6-35
ALTERNATE PROTOCOL: AFFINITY CHROMATOGRAPHY PURIFICATION OF GST FUSION
PROTEIN FROM INCLUSION BODIES 6-36 BASIC PROTOCOL 3: PROTEASE CLEAVAGE
OF FUSION PROTEIN IN SOLUTION TO REMOVE GST AFFINITY TAG 6-37 6.7
EXPRESSION AND PURIFICATION OF THIOREDOXIN FUSION PROTEINS 6-38 BASIC
PROTOCOL: CONSTRUCTION AND EXPRESSION OF A THIOREDOXIN FUSION PROTEIN
6-39 SUPPORT PROTOCOL: OSMOTIC RELEASE OF THIOREDOXIN FUSION PROTEINS
6-41 7 CHARACTERIZATION OF RECOMBINANT PROTEINS 7-1 7.1 OVERVIEW OF THE
CHARACTERIZATION OF RECOMBINANT PROTEINS *** 7-3 7.2 DETERMINING THE
IDENTITY AND PURITY OF RECOMBINANT PROTEINS BY UV ABSORPTION
SPECTROSCOPY 7-7 BASIC PROTOCOL: ANALYSIS OF PROTEINS USING NEAR-UV
SPECTROPHOTOMETRY 7-7 7.3 DETERMINING THE IDENTITY AND STRUCTURE OF
RECOMBINANT PROTEINS 7-10 BASIC PROTOCOL: DETERMINING THE DISULFIDE
LINKAGE PATTERN OF A RECOMBINANT PROTEIN 7-10 7.4 TRANSVERSE
UREA-GRADIENT GEL ELECTROPHORESIS 7-13 7.5 ANALYTICAL
ULTRACENTRIFUGATION 7-17 7.6 DETERMINING THE CD SPECTRUM OF A PROTEIN
7-17 BASIC PROTOCOL: RECORDING A CD SPECTRUM 7-20 SUPPORT PROTOCOL:
INTERPRETATION OF FAR-UV CD SPECTRA 7-22 7.7 DETERMINING THE
FLUORESCENCE SPECTRUM OF A PROTEIN 7-23 BASIC PROTOCOL: RECORDING A
FLUORESCENCE EMISSION SPECTRUM 7-24 7.8 MEASURING PROTEIN
THERMOSTABILITY BY DIFFERENTIAL SCANNING CALORIMETRY 7-27 BASIC
PROTOCOL: DIFFERENTIAL SCANNING CALORIMETRY 7-27 SUPPORT PROTOCOL 1:
ELECTRICAL CALIBRATION 7-28 SUPPORT PROTOCOL 2: TEMPERATURE CALIBRATION
USING HYDROCARBON CAPSULES 7-28 SUPPORT PROTOCOL 3: TEMPERATURE
CALIBRATION USING LIPID SUSPENSIONS 7-29 SUPPORT PROTOCOL 4: MAINTENANCE
AND CLEANING OF DSC CELLS 7-29 7.9 CHARACTERIZING RECOMBINANT PROTEINS
USING HPLC GEL FILTRATION AND MASS SPECTROMETRY 7-30 STRATEGIC PLANNING
7-30 BASIC PROTOCOL: HPLC ANALYSIS OF RECOMBINANT PROTEINS 7-31 SUPPORT
PROTOCOL: MALDI-MS ANALYSIS OF RECOMBINANT PROTEINS 7-33 8 CONVENTIONAL
CHROMATOGRAPHIC SEPARATIONS 8-1 8.1 OVERVIEW OF CONVENTIONAL
CHROMATOGRAPHY 8-1 YIELD AND PURITY OF TARGET PROTEIN 8-1 STEPS IN A
PURIFICATION STRATEGY 8-3 PARAMETERS AFFECTING CHROMATOGRAPHIC
RESOLUTION 8-5 8.2 ION-EXCHANGE CHROMATOGRAPHY 8-10 STRATEGIC PLANNING
8-10 BASIC PROTOCOL 1: BATCH ADSORPTION AND STEP-GRADIENT ELUTION WITH
INCREASING SALT CONCENTRATION - 8-14 ALTERNATE PROTOCOL: PH-BASED
STEP-GRADIENT ELUTION 8-16 BASIC PROTOCOL 2: COLUMN CHROMATOGRAPHY WITH
LINEAR GRADIENT ELUTION 8-16 CONTENTS SHORT PROTOCOLS IN PROTEIN SCIENCE
VII SUPPORT PROTOCOL 1: TEST TUBE PILOT EXPERIMENT TO DETERMINE STARTING
CONDITIONS FOR ION-EXCHANGE CHROMATOGRAPHY 8-17 SUPPORT PROTOCOL 2:
MEASUREMENT OF DYNAMIC (COLUMN) CAPACITY AND BREAKTHROUGH CAPACITY OF
ION-EXCHANGE COLUMNS 8-20 SUPPORT PROTOCOL 3: GRADIENT-FORMATION
TECHNIQUES 8-20 SUPPORT PROTOCOL 4: CLEANING AND REGENERATION OF
ION-EXCHANGE MEDIA 8-21 - SUPPORT PROTOCOL 5: STORAGE OF ION-EXCHANGE
MEDIA 8-21 8.3 GEL-FILTRATION CHROMATOGRAPHY 8-22 STRATEGIC PLANNING
8-22 BASIC PROTOCOL 1: DESALTING (GROUP SEPARATION) 8-23 BASIC PROTOCOL
2: PROTEIN FRACTIONATION 8-27 BASIC PROTOCOL 3: DETERMINATION OF
MOLECULAR SIZE 8-28 SUPPORT PROTOCOL: COLUMN CALIBRATION 8-28 8.4
HYDROPHOBIC-INTERACTION CHROMATOGRAPHY 8-29 STRATEGIC PLANNING 8-29
BASIC PROTOCOL 1: PACKING A COLUMN WITH AN HIC MATRIX OF 90-(IM BEADS
8-31 ALTERNATE PROTOCOL: PACKING A COLUMN WITH AN HIC MATRIX OF 34-JXM
BEADS 8-32 BASIC PROTOCOL 2: TESTING THE PACKED BED * 8-33 BASIC
PROTOCOL 3: ELUTION OF PROTEINS FROM HIC COLUMNS ^ 8-34 SUPPORT
PROTOCOL: REGENERATION, CLEANING, AND STORAGE OF HIC COLUMNS 8-35 8.5
HPLC OF PEPTIDES AND PROTEINS 8-36 START-UP PROCEDURES * 8-36 STANDARD
OPERATING CONDITIONS FOR HP-SEC 8-41 STANDARD OPERATING CONDITIONS FOR
HP-NPC 8-42 STANDARD OPERATING CONDITIONS FOR HP-HIC 8-42 STANDARD
OPERATING CONDITIONS FOR HP-IEX - 8-43 STANDARD OPERATING CONDITIONS FOR
HP-HILIC R 8-43 STANDARD OPERATING CONDITIONS FOR HP-IMAC 8-44 STANDARD
OPERATING CONDITIONS FOR HP-BAC R . 8-45 STANDARD OPERATING CONDITIONS
FOR RP-HPLC 8-45 DESALTING OF PEPTIDE AND PROTEIN MIXTURES BY RP-HPLC
TECHNIQUES 8-47 METHOD DEVELOPMENT IN RP-HPLC 8-47 9 AFFINITY
PURIFICATION 9-1 9.1 LECTIN AFFINITY CHROMATOGRAPHY , 9-3 BASIC
PROTOCOL: CON A-SEPHAROSE AFFINITY CHROMATOGRAPHY 9-3 SUPPORT PROTOCOL:
PILOT STUDY TO DETERMINE LECTIN BINDING AND ELUTION CONDITIONS 9-6
ALTERNATE PROTOCOL: WHEAT GERM AGGLUTININ (WGA)-AGAROSE AFFINITY
CHROMATOGRAPHY 9-6 9.2 DYE AFFINITY CHROMATOGRAPHY 9-7 BASIC PROTOCOL 1:
SELECTION OF COMPONENTS USING CHROMATOGRAPHY 9-7 BASIC PROTOCOL 2:
NEGATIVE CHROMATOGRAPHY 9-9 BASIC PROTOCOL 3: POSITIVE CHROMATOGRAPHY
USING BATCH ELUTION 9-10 SUPPORT PROTOCOL: IMMOBILIZATION OF REACTIVE
DYES 9-10 9.3 AFFINITY PURIFICATION OF NATURAL LIGANDS 9-12 BASIC
PROTOCOL: CYANOGEN BROMIDE ACTIVATION 9-12 ALTERNATE PROTOCOL 1:
ACTIVATION WITH P-NITROPHENYL CHLOROFORMATE 9-15 ALTERNATE PROTOCOL 2:
ACTIVATION WITH TRESYL CHLORIDE 9-16 9.4 METAL-CHELATE AFFINITY
CHROMATOGRAPHY (MCAC) 9-16 STRATEGIC PLANNING - 9-17 BASIC PROTOCOL:
NATIVE MCAC FOR PURIFICATION OF SOLUBLE HISTIDINE-TAIL FUSION PROTEINS
9-17 ALTERNATE PROTOCOL 1: DENATURING MCAC FOR PURIFICATION OF INSOLUBLE
HISTIDINE-TAIL FUSION PROTEINS 9-19 ALTERNATE PROTOCOL 2: SOLID-PHASE
RENATURATION OF MCAC-PURIFIED PROTEINS 9-20 SUPPORT PROTOCOL 1: ANALYSIS
AND PROCESSING OF PURIFIED PROTEINS 9-20 SUPPORT PROTOCOL 2: NTA RESIN
REGENERATION 9-21 SHORT PROTOCOLS IN PROTEIN SCIENCE CONTENTS VIII 9.5
IMMUNOAFFINITY CHROMATOGRAPHY BASIC PROTOCOL: ISOLATION OF SOLUBLE OR
MEMBRANE-BOUND ANTIGENS ALTERNATE PROTOCOL 1: LOW-PH ELUTION OF ANTIGENS
ALTERNATE PROTOCOL 2: BATCH PURIFICATION OF ANTIGENS ALTERNATE PROTOCOL
3: ELUTION IN OCTYL FI-D-GLUCOSIDE SUPPORT PROTOCOL: PREPARATION OF
ANTIBODY-SEPHAROSE 9.6 IMMUNOPRECIPITATION BASIC PROTOCOL 1:
IMMUNOPRECIPITATION USING CELLS IN SUSPENSION LYSED WITH A NONDENATURING
DETERGENT SOLUTION BASIC PROTOCOL 2: IMMUNOPRECIPITATION-RECAPTURE 10
ELECTROPHORESIS 10.1 ONE-DIMENSIONAL SDS GEL ELECTROPHORESIS OF PROTEINS
ELECTRICITY AND ELECTROPHORESIS BASIC PROTOCOL: DENATURING (SDS)
DISCONTINUOUS PAGE: LAEMMLI METHOD ALTERNATE PROTOCOL 1: ELECTROPHORESIS
IN TRIS-TRICINE BUFFER SYSTEMS ALTERNATE PROTOCOL 2: SEPARATION OF
PROTEINS ON GRADIENT GELS SUPPORT PROTOCOL: CASTING MULTIPLE GRADIENT
GELS 10.2 ONE-DIMENSIONAL ELECTROPHORESIS USING NONDENATURING CONDITIONS
BASIC PROTOCOL: CONTINUOUS ELECTROPHORESIS IN NONDENATURING
POLYACRYLAMIDE GELS ALTERNATE PROTOCOL: NATIVE DISCONTINUOUS
ELECTROPHORESIS AND GENERATION OF MOLECULAR WEIGHT STANDARD CURVES
(FERGUSON PLOTS) 10.3 TWO-DIMENSIONAL GEL ELECTROPHORESIS BASIC PROTOCOL
1: ISOELECTRIC FOCUSING USING IMMOBILIZED PH GRADIENT GEL STRIPS BASIC
PROTOCOL 2: SECOND-DIMENSION ELECTROPHORESIS OF IPG GELS ALTERNATE
PROTOCOL: DIAGONAL GEL ELECTROPHORESIS (NONREDUCING/REDUCING GELS) 10.4
PROTEIN DETECTION IN GELS USING FIXATION BASIC PROTOCOL 1: COOMASSIE
BLUE R-250 STAINING ALTERNATE PROTOCOL 1: RAPID COOMASSIE BLUE G-250
STAINING ALTERNATE PROTOCOL 2: ACID-BASED COOMASSIE BLUE G-250 STAINING
BASIC PROTOCOL 2: SYPRO RUBY STAINING BASIC PROTOCOL 3: SILVER STAINING
ALTERNATE PROTOCOL 3: NONAMMONIACAL SILVER STAINING ALTERNATE PROTOCOL
4: RAPID SILVER STAINING SUPPORT PROTOCOL: GEL IMAGING 10.5
ELECTROBLOTTING FROM POLYACRYLAMIDE GELS BASIC PROTOCOL: ELECTROBLOTTING
ONTO PVDF MEMBRANES ALTERNATE PROTOCOL 1: ELECTROBLOTTING OF PROTEINS
FOR SEQUENCE ANALYSIS ALTERNATE PROTOCOL 2: ELECTROBLOTTING ONTO
NITROCELLULOSE MEMBRANES ALTERNATE PROTOCOL 3: PROTEIN ELECTROBLOTTING
IN SEMIDRY SYSTEMS 10.6 DETECTION OF PROTEINS ON BLOT MEMBRANES BASIC
PROTOCOL 1: AMIDO BLACK STAINING BASIC PROTOCOL 2: COOMASSIE BLUE R-250
STAINING BASIC PROTOCOL 3: PONCEAU S STAINING BASIC PROTOCOL 4:
COLLOIDAL GOLD STAINING BASIC PROTOCOL 5: COLLOIDAL SILVER STAINING
BASIC PROTOCOL 6: INDIA INK STAINING BASIC PROTOCOL 7: FLUORESCAMINE
LABELING 10.7 IMMUNOBLOT DETECTION BASIC PROTOCOL: IMMUNOPROBING WITH
DIRECTLY CONJUGATED SECONDARY ANTIBODY ALTERNATE PROTOCOL: IMMUNOPROBING
WITH AVIDIN-BIOTIN COUPLING TO SECONDARY ANTIBODY SUPPORT PROTOCOL 1:
VISUALIZATION WITH CHROMOGENIC SUBSTRATES SUPPORT PROTOCOL 2:
VISUALIZATION WITH LUMINESCENT SUBSTRATES 9-21 9-21 9-24 9-24 9-25 9-25
9-26 9-27 9-33 10-1 10-2 10-2 10-4 10-7 10-9 10-11 10-14 10-14 10-17
10-19 10-19 10-22 10-24 10-25 10-25 10-27 10-28 10-28 10-29 10-30 10-31
10-32 10-34 10-34 10-36 10-37 10-37 10-39 10-39 10-40 10-40 10-41 10-42
10-42 10-43 10-43 10-44 10-45 10-46 10-47 CONTENTS SHORT PROTOCOLS IN
PROTEIN SCIENCE IX 11 CHEMICAL ANALYSIS 11-1 11.1 ENZYMATIC DIGESTION OF
PROTEINS IN SOLUTION 11-2 BASIC PROTOCOL: DIGESTION OF PROTEINS UNDER
NONDENATURING CONDITIONS 11-2 ALTERNATE PROTOCOL 1: DIGESTION OF
PROTEINS IN THE PRESENCE OF UREA OR GUANIDINEHCL 11-4 ALTERNATE PROTOCOL
2: DIGESTION OF PROTEINS IN THE PRESENCE OF SDS 11-5 SUPPORT PROTOCOL 1:
PREPARING AND USING ENZYME STOCKS 11-6 SUPPORT PROTOCOL 2: REDUCTION AND
S-ALKYLATION OF PEPTIDES IN DIGEST MIXTURES 11-9 11.2 ENZYMATIC
DIGESTION OF PROTEINS ON PVDF MEMBRANES 11-10 BASIC PROTOCOL: DIGESTION
OF PVDF-BOUND PROTEINS IN A HYDROGENATED TRITON X-100 BUFFER 11-10 11.3
DIGESTION OF PROTEINS IN GELS FOR SEQUENCE ANALYSIS 11-12 BASIC
PROTOCOL: DIGESTION OF PROTEINS IN GELS IN THE PRESENCE OF TWEEN 20
11-12 ALTERNATE PROTOCOL 1: DIGESTION OF PROTEINS IN GELS IN THE
PRESENCE OF SODIUM DODECYL SULFATE 11-14 ALTERNATE PROTOCOL 2: DIGESTION
OF PROTEINS IN GELS IN THE ABSENCE OF DETERGENT 11-15 11.4 CHEMICAL
CLEAVAGE OF PROTEINS IN SOLUTION 11-15 BASIC PROTOCOL 1: CLEAVAGE AT THE
C TERMINUS OF MET RESIDUES WITH CNBR 11-16 BASIC PROTOCOL 2: CLEAVAGE AT
THE C TERMINUS OF TRP RESIDUES WITH BNPS-SKATOLE 11-17 BASIC PROTOCOL 3:
CLEAVAGE AT ASP-PRO PEPTIDE BONDS WITH FORMIC ACID 11-18 BASIC PROTOCOL
4: CLEAVAGE AT ASN-GLY PEPJIDE BONDS WITH HYDROXYLAMINE 11-18 BASIC
PROTOCOL 5: CLEAVAGE AT THE N TERMINUS OF CYS RESIDUES WITH NTCB 11-18
11.5 CHEMICAL CLEAVAGE OF PROTEINS ON MEMBRANES , 11-20 BASIC PROTOCOL
1: CLEAVAGE AT THE C TERMINUS OF MET RESIDUES WITH CNBR 11-20 ALTERNATE
PROTOCOL: CNBR CLEAVAGE OF PVDF-BOUND PROTEIN PREVIOUSLY ANALYZED BY
EDMAN DEGRADATION 11-21 BASIC PROTOCOL 2: CLEAVAGE AT THE C TERMINUS
OF TRP RESIDUES WITH BNPS_-SKATOLE . 11-21 BASIC PROTOCOL 3: CLEAVAGE AT
ASP-PRO PEPTIDE BONDS WITH FORMIC ACID 11-22 BASIC PROTOCOL 4: CLEAVAGE
AT ASN-GLY PEPTIDE BONDS WITH HYDROXYLAMINE 11-23 BASIC PROTOCOL 5:
CLEAVAGE AT THE N TERMINUS OF CYS RESIDUES WITH NTCB 11-23 11.6
REVERSED-PHASE ISOLATION OF PEPTIDES 11-24 BASIC PROTOCOL 1:
REVERSED-PHASE PEPTIDE SEPARATION AT THE 5- TO 500-PMOL LEVEL 11-24
BASIC PROTOCOL 2: REVERSED-PHASE PEPTIDE SEPARATION AT 5 PMOL 11-26
CAPILLARY HPLC SYSTEM ASSEMBLY 11-26 11.7 N-TERMINAL SEQUENCE ANALYSIS
11-28 INSTRUMENTATION 11-28 SAMPLE PREPARATION 11-29 12
POST-TRANSLATIONAL MODIFICATION: GLYCOSYLATION 12-1 12.1 INHIBITION OF
N-LINKED GLYCOSYLATION 12-1 BASIC PROTOCOL: INHIBITION OF N-LINKED
GLYCOSYLATION 12-1 SUPPORT PROTOCOL: ACETONE PRECIPITATION 12-5 12.2
ENDOGLYCOSIDASE AND GLYCOAMIDASE RELEASE OF N-LINKED OLIGOSACCHARIDES
12-6 BASIC PROTOCOL 1: ENDOGLYCOSIDASE H DIGESTION 12-7 BASIC PROTOCOL
2: PEPTIDE:N-GLYCOSIDASE F DIGESTION 12-7 SUPPORT PROTOCOL: ESTIMATING
THE NUMBER OF N-LINKED OLIGOSACCHARIDE CHAINS ON A GLYCOPROTEIN 12-8
BASIC PROTOCOL-3: SIALIDASE (NEURAMINIDASE) DIGESTION - 12-9 12.3
DETECTION OF GLYCOPHOSPHOLIPID ANCHORS ON PROTEINS 12-9 BASIC PROTOCOL
1: EXTRACTION AND PARTITIONING OF TOTAL PROTEINS FROM CELLS OR MEMBRANES
WITH TRITON X-L 14 12-10 ALTERNATE PROTOCOL: PARTITIONING OF ISOLATED
PROTEINS WITH TRITON X-L 14 12-10 BASIC PROTOCOL 2: IDENTIFICATION OF
GPI-ANCHORED PROTEINS BY PI-PLC DIGESTION OF INTACT CELLS 12-10 SHORT
PROTOCOLS IN PROTEIN SCIENCE CONTENTS 13 POST-TRANSLATIONAL
MODIFICATION: PHOSPHORYLATION AND PHOSPHATASES 13-1 13.1 LABELING
CULTURED CELLS WITH 32 P. AND PREPARING CELL LYSATES FOR
IMMUNOPRECIPITATION 13-2 BASIC PROTOCOL: LABELING CULTURED CELLS WITH
32 P AND LYSIS USING MILD DETERGENT 13-2 ALTERNATE PROTOCOL: LYSIS OF
CELLS BY BDILING IN SDS . , Y 13-3 13.2 PHOSPHOAMINO ACID ANALYSIS 13-4
BASIC PROTOCOL: ACID HYDROLYSIS AND TWO-DIMENSIONAL ELECTROPHORETIC
ANALYSIS OF PHOSPHOAMINO ACIDS 13-4 ALTERNATE PROTOCOL: ALKALI TREATMENT
TO ENHANCE DETECTION OF TYR- AND THR-PHOSPHORYLATED PROTEINS BLOTTED
ONTO FILTERS 13-7 13.3 DETECTION OF PHOSPHORYLATION BY IMMUNOLOGICAL
TECHNIQUES 13-9 BASIC PROTOCOL: IMMUNOBLOTTING WITH ANTI-PHOSPHOTYROSINE
ANTIBODIES AND DETECTION USING [ 125 I]PROTEIN A 13-9 ALTERNATE
PROTOCOL: DETECTION OF BOUND ANTIBODIES BY ENHANCED CHEMILUMINESCENCE
(ECL) 13-10 13.4 DETECTION OF PHOSPHORYLATION BY ENZYMATIC TECHNIQUES
13-11 BASIC PROTOCOL 1: DIGESTION OF PHOSPHOPROTEINS WITH NONSPECIFIC
ACID PHOSPHATASES 13-11 ALTERNATE PROTOCOL 1: DIGESTION OF
PHOSPHOPROTEINS WITH NONSPECIFIC ALKALINE PHDSPHATASE 13-12 BASIC
PROTOCOL 2: DIGESTION OF PHOSPHOPROTEINS WITH PROTEIN SERINE/THREONINE
PHOSPHATASES 13-13 ALTERNATE PROTOCOL 2: DIGESTION OF PHOSPHOPROTEINS
WITH PROTEIN TYROSINE PHOSPHATASES 13-13 13.5 PERMEABILIZATION
STRATEGIES TO STUDY PROTEIN PHOSPHORYLATION 13-14 BASIC PROTOCOL:
ANALYSIS OF PROTEIN PHOSPHORYLATION IN PERMEABILIZED CELLS ** S 13-14
13.6 PHOSPHOPEPTIDE MAPPING AND IDENTIFICATION OF PHOSPHORYLATION SITES
13-16 BASIC PROTOCOL: TRYPTIC PHOSPHOPEPTIDE MAPPING OF PROTEINS
ISOLATED FROM SDS-POLYACRYLAMIDE GELS 13-16 ALTERNATE PROTOCOL:
PROTEOLYTIC DIGESTION OF IMMOBILIZED PROTEINS 13-22 SUPPORT PROTOCOL:
PREPARATION OF PHOSPHOPEPTIDES FOR MICROSEQUENCE DETERMINATION OR MASS
SPECTROMETRY 13-22 14 POST-TRANSLATIONAL MODIFICATION: SPECIALIZED
APPLICATIONS 14-1 14.1 ANALYSIS OF DISULFIDE BOND FORMATION 14-2 BASIC
PROTOCOL: ANALYSIS OF DISULFIDE BOND FORMATION IN INTACT MONOLAYER CELLS
14-2 ALTERNATE PROTOCOL: ANALYSIS OF DISULFIDE BOND FORMATION IN CELLS
IN SUSPENSION 14-3 14.2 ANALYSIS OF PROTEIN ACYLATION - 14-4 BASIC
PROTOCOL 1: BIOSYNTHETIC LABELING WITH FATTY ACIDS 14-4 BASIC PROTOCOL
2: ANALYSIS OF FATTY ACID LINKAGE TO PROTEIN 14-5 BASIC PROTOCOL 3:
ANALYSIS OF TOTAL PROTEIN-BOUND FATTY ACID LABEL IN CELL EXTRACT 14-6
BASIC PROTOCOL 4: ANALYSIS OF FATTY ACID LABEL IDENTITY 14-7 14.3
ANALYSIS OF PROTEIN PRENYLATION AND CARBOXYL-METHYLATION 14-7 BASIC
PROTOCOL 1: PRENYLATION OF PROTEINS IN CULTURED CELLS - 14-7 BASIC
PROTOCOL 2: CARBOXYL-METHYLATION OF PROTEINS IN CULTURED CELLS 14-8
14.4 ANALYSIS OF OXIDATIVE MODIFICATION OF PROTEINS 14-10 BASIC PROTOCOL
1: SPECTROPHOTOMETRIC QUANTITATION OF PROTEIN CARBONYLS USING
2,4-DINITROPHENYLHYDRAZINE 14-10 BASIC PROTOCOL 2: QUANTITATION OF
PROTEIN CARBONYLS DERIVATIZED WITH TRITIATED SODIUM BOROHYDRIDE 14-11
BASIC PROTOCOL 3: GEL ELECTROPHORETIC ANALYSIS OF PROTEIN THIOL GROUPS
LABELED WITH [ 14 C]IODOACETAMIDE 14-12 BASIC PROTOCOL 4:
QUANTIFICATION OF PROTEIN DITYROSINE RESIDUESBY MASS SPECTROMETRY 14-12
SUPPORT PROTOCOL 1: PREPARATION OF O,O -DITYROSINE STANDARD 14-14
SUPPORT PROTOCOL 2: ANALYSIS OF PROTEIN-BOUND NITROTYROSINE BY A
COMPETITIVE ELIS A METHOD 14-15 BASIC PROTOCOL 5: ENZYMATIC ANALYSIS OF
ISOASPARTATE FORMATION 14-16 CONTENTS SHORT PROTOCOLS IN PROTEIN SCIENCE
XI 14.5 ANALYSIS OF PROTEIN UBIQUITINATION / BASIC PROTOCOL 1:
IMMUNOPRECIPITATION OF TARGET PROTEIN FOLLOWED BY ANTI-UB IMMUNOBLOTTING
BASIC PROTOCOL 2: AFFINITY PURIFICATION OF UBIQUITINATED PROTEINS FROM
CELLS EXPRESSING HIS 6 -UB 15 CHEMICAL MODIFICATION OF PROTEINS 15.1
MODIFICATION OF CYSTEINE STRATEGIC PLANNING BASIC PROTOCOL 1: ALKYLATION
OF A PROTEIN OF KNOWN SIZE AND COMPOSITION WITH HALOAC YL REAGENTS OR
,/V-EFHYLMALEIMIDE BASIC PROTOCOL 2: ALKYLATION WITH
^-(IODOEFHYLJ-TRIFLUOROACETAMIDE BASIC PROTOCOL 3: ALKYLATION WITH
ACRYLAMIDE BASIC PROTOCOL 4: AIR OXIDATION TO A DISULFIDE SUPPORT
PROTOCOL 1: COLORIMETRIC QUANTITATION OF FREE SULFHYDRYLS SUPPORT
PROTOCOL 2: DESALTING THE SAMPLE AFTER CYSTEINE MODIFICATION 15.2
MODIFICATION OF AMINO GROUPS STRATEGIC PLANNING BASIC PROTOCOL 1:
AMIDATION USING A SUCCINIMIDYL ESTER BASIC PROTOCOL 2: ADDITION OF
FLUORESCEIN ISOTHIOCYANATE BASIC PROTOCOL 3: SUCCINYLATION BASIC
PROTOCOL 4: REDUCTIVE METHYLATION / - 16 MASS SPECTROMETRY 16.1
OVERVIEW OF PEPTIDE AND PROTEIN ANALYSIS BY MASS SPECTROMETRY WHY IS MS
AN ESSENTIAL TOOL IN PROTEIN STRUCTURE ANALYSIS? WHAT IS MS? WHAT IS
TANDEM MS? IS MS DATA QUANTITATIVE? SAMPLE PREPARATION FUNDAMENTALS OF
MASS MEASUREMENT ACCURACY AND MASS RESOLUTION 16.2 SAMPLE PREPARATION
FOR MALDI MASS ANALYSIS OF PEPTIDES AND PROTEINS BASIC PROTOCOL 1: DRIED
DROPLET METHOD BASIC PROTOCOL 2: RAPID EVAPORATION METHOD SUPPORT
PROTOCOL: PURIFICATION/RECRYSTALLIZATION OF HCCA 16.3 IN-GEL DIGESTION
OF PROTEINS FOR MALDI-MS FINGERPRINT MAPPING 16.4 SEARCHING SEQUENCE
DATABASES OVER THE INTERNET: PROTEIN IDENTIFICATION USING MS-FIT 16.5
SEARCHING SEQUENCE DATABASES OVER THE INTERNET: PROTEIN IDENTIFICATION
USING MS-TAG 16.6 INTRODUCING SAMPLES DIRECTLY INTO ELECTROSPRAY
IONIZATION MASS SPECTROMETERS USING A NANOSPRAY INTERFACE 16.7
INTRODUCING SAMPLES DIRECTLY INTO ELECTROSPRAY IONIZATION MASS
SPECTROMETERS USING MICROSCALE CAPILLARY LIQUID CHROMATOGRAPHY BASIC
PROTOCOL 1: BUILDING AN INTEGRATED LC COLUMN ES NEEDLE BASIC PROTOCOL 2:
PACKING THE ES NEEDLE BASIC PROTOCOL 3: MOUNTING AND USING THE
MICROSCALE ES LC/MS INTERFACE ASSEMBLY 16.8 PROTEIN IDENTIFICATION USING
A QUADRUPOLE ION TRAP MASS SPECTROMETER AND SEQUEST DATABASE MATCHING
16.9 DE NOVO PEPTIDE SEQUENCING VIA MANUAL INTERPRETATION OF MS/MS
SPECTRA BASIC PROTOCOL: MANUAL INTERPRETATION OF MS/MS SPECTRA SUPPORT
PROTOCOL 1: CONFIRMATION OF THE DE NOVO SEQUENCE THROUGH METHYL
ESTERIFICATION SUPPORT PROTOCOL 2: CONFIRMATION OF THE DE NOVO SEQUENCE
THROUGH ACETYLATION 14-17 14-17 14-18 15-1 15-2 15-2 15-3 15-4 15-5 15-5
15-6 15-7 15-8 15-8 15-8 - 15-9 15-12 15-12 16-1 16-2 16-2 16-3 16-4
16-5 16-5 16-6 16-8 16-10 16-10 16-11 16-12 16-13 16-14 16-15 16-17
16-17 16-18 16-19 16-20 16-21 16-21 16-26 16-27 SHORT PROTOCOLS IN
PROTEIN SCIENCE CONTENTS XII 17 PREPARATION AND HANDLING OF PEPTIDES
17-1 17.1 SYNTHESIS OF MULTIPLE PEPTIDES ON PLASTIC PINS 17-1 BASIC
PROTOCOL: MULTIPIN SYNTHESIS OF PEPTIDES J 17-3 SUPPORT PROTOCOL 1:
PREPARING ACTIVATED FMOC-PROTECTED AMINO ACID SOLUTIONS - 17-5 SUPPORT
PROTOCOL 2: N-TERMINAL ACETYLATION OF PEPTIDES 17-6 SUPPORT PROTOCOL 3:
N-TERMINAL BIOTINYLATIONOF PEPTIDES 17-6 17.2 SYNTHETIC PEPTIDES FOR
PRODUCTION OF ANTIBODIES THAT RECOGNIZE INTACT PROTEINS 17-7 BASIC
PROTOCOL 1: COMPUTER-ASSISTED SELECTION OF APPROPRIATE ANTIGENIC PEPTIDE
SEQUENCES 17-7 ALTERNATE PROTOCOL 1: MANUAL INSPECTION TO SELECT
APPROPRIATE PEPTIDE SEQUENCES 17-8 BASIC PROTOCOL 2: DESIGNING A
SYNTHETIC PEPTIDE FOR COUPLING TO A CARRIER PROTEIN 17-9 ALTERNATE
PROTOCOL 2: DESIGNING A SYNTHETIC MULTIPLE ANTIGENIC PEPTIDE 17-10 BASIC
PROTOCOL 3: COUPLING SYNTHETIC PEPTIDES TO A CARRIER PROTEIN USING A
HETEROBIFUNCTIONAL REAGENT 17-10 SUPPORT PROTOCOL 1: ASSAY OF FREE
SULFHYDRYLS WITHEUMAN S REAGENT 17-11 SUPPORT PROTOCOL 2: REDUCING
CYSTEINE GROUPS IN PEPTIDES 17-11 ALTERNATE PROTOCOL 3: COUPLING
SYNTHETIC PEPTIDES TO A CARRIER PROTEIN USING A HOMOBIFUNCTIONAL REAGENT
17-13 ALTERNATE PROTOCOL 4: COUPLING SYNTHETIC PEPTIDES TO A CARRIER
PROTEIN USING A CARBODIIMIDE 17-13 SUPPORT PROTOCOL 3: CALCULATION OF
THE MOLAR RATIO OF PEPTIDE TO CARRIER PROTEIN 17-14 17.3 SYNTHESIS AND
APPLICATION OF PEPTIDE DENDRIMERS AS PROTEIN MIMETICS 17-15 BASIC
PROTOCOL 1: DIRECT BOC SYNTHESIS OF MAP SYSTEMS S 17-17 ALTERNATE
PROTOCOL: DIRECT FMOC SOLID-PHASE SYNTHESIS OF MAP SYSTEMS 17-19 SUPPORT
PROTOCOL L:NINHYDRIN TEST ~^7 17-21 SUPPORT PROTOCOL 2: PURIFICATION OF
MAP SYSTEM BY DIALYSIS 17-21 SUPPORT PROTOCOL 3: PURIFICATION OF MAP
USING HIGH-PERFORMANCE GEL-FILTRATION CHROMATOGRAPHY 17-22 BASIC
PROTOCOL 2: DIRECT SYNTHESIS OF END-TO-SIDE CHAIN CMAP 17-22 17.4
DISULFIDE BOND FORMATION IN PEPTIDES 17-24 BASIC PROTOCOL 1: PROMOTING
DISULFIDE BOND FORMATION BY AIR OXIDATION 17-24 ALTERNATE PROTOCOL 1:
PROMOTING DISULFIDE BOND FORMATION BY ON-RESIN AIR OXIDATION 17-24
ALTERNATE PROTOCOL 2: CHARCOAL/AIR-MEDIATED INTRAMOLECULAR DISULFIDE
FORMATION Y 17-25 BASIC PROTOCOL 2: INTRAMOLECULAR DISULFIDE FORMATION
BY POTASSIUM FERRICYANIDE OXIDATION 17-25 ALTERNATE PROTOCOL 3:
INTERMOLECULAR OR INTRAMOLECULAR DISULFIDE FORMATION BY POTASSIUM
FERRICYANIDE OXIDATION 17-26 ALTERNATE PROTOCOL 4: ON-RESIN DISULFIDE
FORMATION BY POTASSIUM FERRICYANIDE OXIDATION 17-26 BASIC PROTOCOL 3:
OXIDATION UNDER SLIGHTLY ACIDIC PH CONDITIONS BY DMSO 17-27 ALTERNATE
PROTOCOL 5: OXIDATION UNDER SLIGHTLY BASIC PH CONDITIONS BY DMSO 17-27
BASIC PROTOCOL 4: OXIDATION BY REDOX BUFFERS ,. 17-28 BASIC PROTOCOL 5:
OXIDATION MEDIATED BY SOLID-PHASE ELLMAN S REAGENT 17-28 BASIC PROTOCOL
6: SIMULTANEOUS DEPROTECTION/OXIDATION WITH IODINE 17-29 ALTERNATE
PROTOCOL 6: SIMULTANEOUS ON-RESIN DEPROTECTION/OXIDATION OF 5-ACM WITH
IODINE 17-30 BASIC PROTOCOL 7: SIMULTANEOUS DEPROTECTION/OXIDATION OF
S-ACM WITHTL(III) 17-30 ALTERNATE PROTOCOL 7: ON-RESIN SIMULTANEOUS
DEPROTECTION/OXIDATION OF 5-ACM WITHTL(III) S . 17-31 BASIC PROTOCOL 8:
ALKYLTRICHLOROSILANE-SULFOXIDE OXIDATION 17-32 18 IDENTIFICATION OF
PROTEIN INTERACTIONS 18-1 18.1 ANALYSIS OF PROTEIN-PROTEIN INTERACTIONS
/ 18-2 BASIC CONCEPTS 18-2 EQUILIBRIUM PARAMETERS 18-2 KINETIC
PARAMETERS 18-4 18.2 INTERACTION TRAP/TWO-HYBRID SYSTEM TO IDENTIFY
INTERACTING PROTEINS 18-5 CONTENTS SHORT PROTOCOLS IN PROTEIN SCIENCE
XIII 18.3 PHAGE-BASED EXPRESSION CLONING TO IDENTIFY INTERACTING
PROTEINS STRATEGIC PLANNING 18.4 DETECTION OF PROTEIN-PROTEIN
INTERACTIONS BY COPRECIPITATION BASIC PROTOCOL: COPRECIPITATING PROTEINS
WITH PROTEIN A- OR PROTEIN G-SEPHAROSE ALTERNATE PROTOCOL:
COPRECIPITATING A GST FUSION PROTEIN 18.5 HIGH-THROUGHPUT SCREENING FOR
PROTEIN-PROTEIN INTERACTIONS USING YEAST TWO-HYBRID ARRAYS BASIC
PROTOCOL 1: CONSTRUCTION OF A PROTEOME-SCALE PROTEIN ARRAY FOR YEAST
BASIC PROTOCOL 2: MANUAL SCREENING FOR PROTEIN INTERACTIONS USING A
YEAST PROTEIN ARRAY 18.6 IDENTIFICATION OF PROTEIN INTERACTIONS BY FAR
WESTERN ANALYSIS BASIC PROTOCOL: FAR WESTERN ANALYSIS OF A PROTEIN
MIXTURE ALTERNATE PROTOCOL 1: DETECTING INTERACTING PROTEINS BY
IMMUNOBLOTTING ALTERNATE PROTOCOL 2: USING PEPTIDES TO IDENTIFY SPECIFIC
INTERACTING SEQUENCES IN A FAR WESTERN BLOT 18.7 SCINTILLATION PROXIMITY
ASSAY (SPA) TECHNOLOGY TO STUDY BIOMOLECULAR INTERACTIONS 19
QUANTITATION OF PROTEIN INTERACTIONS 19.1 OVERVIEW OF THE QUANTITATION
OF PROTEIN INTERACTIONS SURFACE PLASMON RESONANCE ANALYTICAL
ULTRACENTRIFUGATION (SEDIMENTATION EQUILIBRIUM AND SEDIMENTATION
VELOCITY) 19.2 TITRATION CALORIMETRY BASIC PROTOCOL 1: DETERMINING A
MOLAR RATIO, OBSERVED AFFINITY (K^*), AND OBSERVED - BINDING ENTHALPY
CHANGE (AH BS ) BY ISOTHERMAL TITRATION CALORIMETRY BASIC PROTOCOL 2:
DETERMINING BIOCHEMICAL BINDING THERMODYNAMICS: AG , AH AND AC PO/ BY
ITC 19.3 REDUCED-SCALE LARGE-ZONE ANALYTICAL GEL-FILTRATION
CHROMATOGRAPHY FOR MEASUREMENT OF PROTEIN ASSOCIATION EQUILIBRIA BASIC
PROTOCOL: REDUCED-SCALE LARGE-ZONE ANALYTICAL GEL-FILTRATION
CHROMATOGRAPHY SUPPORT PROTOCOL 1: DATA ANALYSIS SUPPORT PROTOCOL 2:
DETERMINATION OF THE ENERGETICS AND STOICHIOMETRY OF THE PROTEIN
ASSEMBLY PROCESS 19.4 SIZE-EXCLUSION CHROMATOGRAPHY WITH ON-LINE LIGHT
SCATTERING BASIC PROTOCOL 1: USING REFRACTIVE INDEX AND LIGHT SCATTERING
TO CALCULATE THE MOLECULAR WEIGHT AND DEGREE OF SELF-ASSOCIATION OF
PROTEINS CONTAINING NO CARBOHYDRATES (TWO-DETECTOR METHOD) ALTERNATE
PROTOCOL 1: COMBINATION OF LIGHT-SCATTERING AND UV DETECTORS TO
CALCULATE THE MOLECULAR WEIGHT OF NONGLYCOSYLATED PROTEINS (TWO-DETECTOR
METHOD) ALTERNATE PROTOCOL 2: SIZE-EXCLUSION CHROMATOGRAPHY WITH LIGHT
SCATTERING FOR LARGE PROTEINS: DEBYE ANALYSIS ALTERNATE PROTOCOL 3:
ABSOLUTE MOLECULAR-WEIGHT CALIBRATION METHOD BASIC PROTOCOL 2:
CALCULATING THE STOICHIOMETRY OF A PROTEIN-PROTEIN COMPLEX CONTAINING NO
CARBOHYDRATES USING LIGHT SCATTERING AND REFRACTIVE INDEX ALTERNATE
PROTOCOL 4: CALCULATING THE STOICHIOMETRY OF PROTEIN-PROTEIN
INTERACTIONS FOR NONGLYCOSYLATED PROTEINS USING LIGHT SCATTERING AND UV
20 PEPTIDASES 20.1 PROTEASES BIOLOGICAL IMPORTANCE OF PROTEASES THE
TERMINOLOGY OF PROTEASES ; THREE WAYS IN WHICH THE MANY PROTEASES MAY
USEFULLY BE SUBDIVIDED 20.2 PURIFICATION AND CHARACTERIZATION OF
PROTEASOMES FROM SACCHAROMYCES CEREVISIAE BASIC PROTOCOL 1: PURIFICATION
OF YEAST 26S PROTEASOME BY ANION-EXCHANGE CHROMATOGRAPHY AND GEL
FILTRATION ALTERNATE PROTOCOL: PURIFICATION OF YEAST 26S PROTEASOME BY
AFFINITY CHROMATOGRAPHY BASIC PROTOCOL 2: PURIFICATION OF YEAST 20S
PROTEASOME BY CONVENTIONAL CHROMATOGRAPHY SHORT PROTOCOLS IN PROTEIN
SCIENCE 18-14 18-14 18-16 18-17 18-18 18-18 18-18 / 18-21 18-22 18-22
18-23 18-24 18-24 19-1 19-2 19-2 19-5 19-6 19-6 19-10 19-12 19-12 19-14
19-15 19-17 19-17 19-18 19-19 19-19 19-20 19-20 20-1 20-1 20-1 20-2 20-2
20-11 20-11 20-12 20-13 CONTENTS XIV SUPPORT PROTOCOL 1: PEPTIDASE
ACTIVITY ASSAY FOR 26S AND 20S PROTEASOME SUPPORT PROTOCOL 2:
PREPARATION OF POLYUBIQUITINATED LYSOZYME SUPPORT PROTOCOL 3:
PREPARATION OF RADIOLABELED CASEIN SUPPORT PROTOCOL 4: DEGRADATION OF
PROTEIN SUBSTRATES BY THE 26S PROTEASOME SUPPORT PROTOCOL 5: NATIVE GEL
ELECTROPHORESIS/IN-GEL PEPTIDASE ASSAY OF THE PROTEASOME 20.3
PURIFICATION OF THE EUKARYOTIC 20S PROTEASOME BASIC PROTOCOL:
PURIFICATION OF THE 20S PROTEASOME FROM BOVINE PITUITARIES SUPPORT
PROTOCOL: ENZYME ASSAY TO ANALYZE PROTEASOME-CATALYZED CLEAVAGE 20.4
SERPINS (SERINE PROTEASE INHIBITORS) BASIC PROTOCOL 1: PURIFICATION OF
HUMAN ANTITHROMBIN BY COLUMN CHROMATOGRAPHY SUPPORT PROTOCOL: ASSAY FOR
ANTITHROMBIN IN ELUTED FRACTIONS BASIC PROTOCOL 2: ASSAY OF ANTITHROMBIN
INHIBITION IN THE ABSENCE OF GLYCOSAMINOGLYCAN* PROGRESSIVE ANTITHROMBIN
ACTIVITY BASIC PROTOCOL 3: AN ASSAY OF ANTITHROMBIN INHIBITION IN THE
PRESENCE OF GLYCOSAMINOGLYCAN* HEPARIN COFACTOR ACTIVITY 20.5 USE OF
GFP AS A REPORTER FOR THE^ANALYSIS OF SEQUENCE-SPECIFIC PROTEASES BASIC
PROTOCOL: FLUORESCENT ASSAY OF SITE-SPECIFIC PROTEASE SUPPORT PROTOCOL:
SUBSTRATE-GFP FUSION PROTEIN PURIFICATION USING METAL-CHELATE
CHROMATOGRAPHY / ALTERNATE PROTOCOL: FLUORESCENT RESONANCE ENERGY
TRANSFER ASSAY OF SITE-SPECIFIC PROTEASES ^ 20.6 OVEREXPRESSION AND
PURIFICATION OF ACTIVE SERINE PROTEASES AND THEIR VARIANTS FROM
ESCHERICHIA COLI INCLUSION BODIES BASIC PROTOCOL 1: OVEREXPRESSION AND
PURIFICATION OF MICROPLASMINOGEN AND ITS VARIANTS FROM ESCHERICHIA COLI
INCLUSION BODIES BASIC PROTOCOL 2: ACTIVATION OF THE ZYMOGEN
MICROPLASMINOGEN AND PURIFICATION OF THE CATALYTICALLY ACTIVE
MICROPLASMIN SUPPORT PROTOCOL 1: ASSAY TO DETERMINE THE ACTIVITY OF THE
RECOMBINANT MICROPLASMIN SUPPORT PROTOCOL 2: ASSAY TO DETERMINE THE
ACTIVE-SITE CONCENTRATION -^ 20.7 ASSAYING PROTEASES IN CELLULAR
ENVIRONMENTS BASIC PROTOCOL 1: MONITORING INTRACELLULAR PROTEASE
ACTIVITY IN SITU WITH-CELL-PERMEABLE PEPTIDE SUBSTRATE BASIC PROTOCOL 2:
MONITORING INTRACELLULAR PROTEASE ACTIVITY IN SITU BY AUTOLYTIC
ACTIVATION AND ENDOGENOUS SUBSTRATE PROTEIN DEGRADATION BASIC PROTOCOL
3: MONITORING PROTEASE ACTIVITY LEVELS AS REFLECTED IN CELL LYSATE BASIC
PROTOCOL 4: MONITORING ACTIVITY OF SECRETED PROTEASE IN SITU BASIC
PROTOCOL 5: USING ZYMOGRAPHY TO MEASURE ACTIVITY OF CELL-SECRETED MMPS
20.8 EXPRESSION, PURIFICATION, AND CHARACTERIZATION OF CASPASES BASIC
PROTOCOL 1: EXPRESSION OF CASPASES IN E. COLI BASIC PROTOCOL 2: CASPASE
ENZYMATIC ASSAY SUPPORT PROTOCOL 1: TITRATION OF RECOMBINANT CASPASE
SUPPORT PROTOCOL 2: PREPARATION OF CASPASE SUBSTRATES 21 GEL-BASED
PROTEOME ANALYSIS 20-14 20-15 20-17 20-18 20-18 20-19 20-19 20-21 20-22
20-22 20-24 20-25 20-26 20-27 20-27 20-28 20-29 20-30 20-30 20-32 20-33
20-33 20-34 20-34 20-35 20-37 20-37 20-38 20-39 20-39 20-43 20-43 20-44
21-1 21.1 PROTEIN PROFILING USING TWO-DIMENSIONAL DIFFERENCE GEL
ELECTROPHORESIS (2-D DIGE) 21.1 BASIC PROTOCOL: LABELING OF PROTEIN WITH
CYANINE DYES (CYDYE DIGE FLUORS) FOR 2-D PAGE 21-1 SUPPORT PROTOCOL:
PREPARATION OF PROTEIN FOR CY LABELING REACTIONS 21-4 ALTERNATE
PROTOCOL: PROTEIN IDENTIFICATION FROM DIGE GELS BY MASS SPECTROMETRIC
TECHNIQUES 21-6 21.2 LASER CAPTURE MICRODISSECTION FOR PROTEOME
ANALYSIS 21.6 BASIC PROTOCOL 1: LASER CAPTURE MICRODISSECTION (LCM) 21-6
ALTERNATE PROTOCOL: IMMUNOLABELING OF TISSUE SECTIONS FOR LCM 21-8
SUPPORT PROTOCOL 1: COLLECTION AND STORAGE OF TISSUE FOR ANALYSIS BY LCM
21-9 SUPPORT PROTOCOL 2: QUANTIFICATION OF PROTEIN IN MATERIAL CAPTURED
BY LCM USING SYPRO RUBY 21-9 BASIC PROTOCOL 2: ANALYSIS OF MATERIAL
CAPTURED BY LCM USING IMMUNOBLOTTING 21-10 BASIC PROTOCOL 3: SELDI MASS
SPECTROMETRY 21-11 CONTENTS SHORT PROTOCOLS IN PROTEIN SCIENCE XV
APPENDICES AL REAGENTS AND SOLUTIONS AL-1 A2 USEFUL MEASUREMENTS AND
DATA A2-1 A3 COMMONLY USED TECHNIQUES A3-1 3 A USE OF PROTEIN FOLDING
REAGENTS A3-1 3B DIALYSIS A3-4 3C SILANIZING GLASSWARE A3-6 3D PROTEIN
PRECIPITATION USING AMMONIUM SULFATE A3-7 3E AGAROSE GEL ELECTROPHORESIS
A3-14 3F QUANTITATION OF NUCLEIC ACIDS WITH ABSORPTION SPECTROSCOPY
A3-16 A4 SELECTED SUPPLIERS OF REAGENTS AND EQUIPMENT A4-1 REFERENCES
INDEX SHORT PROTOCOLS IN PROTEIN SCIENCE CONTENTS XVI
|
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| callnumber-first | Q - Science |
| callnumber-label | QP551 |
| callnumber-raw | QP551 |
| callnumber-search | QP551 |
| callnumber-sort | QP 3551 |
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| classification_rvk | WC 4170 |
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| dewey-ones | 572 - Biochemistry |
| dewey-raw | 572/.6 |
| dewey-search | 572/.6 |
| dewey-sort | 3572 16 |
| dewey-tens | 570 - Biology |
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| id | DE-604.BV019354214 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T19:58:21Z |
| institution | BVB |
| isbn | 0471483389 |
| language | English |
| lccn | 2003012325 |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-012818185 |
| oclc_num | 52381115 |
| open_access_boolean | |
| owner | DE-19 DE-BY-UBM DE-92 DE-355 DE-BY-UBR |
| owner_facet | DE-19 DE-BY-UBM DE-92 DE-355 DE-BY-UBR |
| physical | Getr. Zählung Ill., graph. Darst. |
| publishDate | 2003 |
| publishDateSearch | 2003 |
| publishDateSort | 2003 |
| publisher | Wiley |
| record_format | marc |
| series2 | Current protocols |
| spelling | Short protocols in protein science a compendium of methods from "Current protocols in protein science" ed. by John E. Coligan ... [Hoboken. N.J.] Wiley 2003 Getr. Zählung Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Current protocols Includes bibliographical references and index Eiwitsynthese gtt Eiwitten gtt Proteínas (análise;isolamento e purificação) larpcal Protéines - Purification - Manuels de laboratoire Scheidingstechnieken gtt Spectrometrie gtt Proteins Purification Laboratory manuals Proteins analysis Laboratory Manuals Proteins isolation & purification Laboratory Manuals Methode (DE-588)4038971-6 gnd rswk-swf Proteine (DE-588)4076388-2 gnd rswk-swf Biochemie (DE-588)4006777-4 gnd rswk-swf Proteine (DE-588)4076388-2 s Biochemie (DE-588)4006777-4 s Methode (DE-588)4038971-6 s DE-604 Coligan, John E. Sonstige oth http://www.loc.gov/catdir/description/wiley0310/2003012325.html Publisher description http://www.loc.gov/catdir/toc/wiley031/2003012325.html Table of contents HEBIS Datenaustausch Darmstadt application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=012818185&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Short protocols in protein science a compendium of methods from "Current protocols in protein science" Eiwitsynthese gtt Eiwitten gtt Proteínas (análise;isolamento e purificação) larpcal Protéines - Purification - Manuels de laboratoire Scheidingstechnieken gtt Spectrometrie gtt Proteins Purification Laboratory manuals Proteins analysis Laboratory Manuals Proteins isolation & purification Laboratory Manuals Methode (DE-588)4038971-6 gnd Proteine (DE-588)4076388-2 gnd Biochemie (DE-588)4006777-4 gnd |
| subject_GND | (DE-588)4038971-6 (DE-588)4076388-2 (DE-588)4006777-4 |
| title | Short protocols in protein science a compendium of methods from "Current protocols in protein science" |
| title_auth | Short protocols in protein science a compendium of methods from "Current protocols in protein science" |
| title_exact_search | Short protocols in protein science a compendium of methods from "Current protocols in protein science" |
| title_full | Short protocols in protein science a compendium of methods from "Current protocols in protein science" ed. by John E. Coligan ... |
| title_fullStr | Short protocols in protein science a compendium of methods from "Current protocols in protein science" ed. by John E. Coligan ... |
| title_full_unstemmed | Short protocols in protein science a compendium of methods from "Current protocols in protein science" ed. by John E. Coligan ... |
| title_short | Short protocols in protein science |
| title_sort | short protocols in protein science a compendium of methods from current protocols in protein science |
| title_sub | a compendium of methods from "Current protocols in protein science" |
| topic | Eiwitsynthese gtt Eiwitten gtt Proteínas (análise;isolamento e purificação) larpcal Protéines - Purification - Manuels de laboratoire Scheidingstechnieken gtt Spectrometrie gtt Proteins Purification Laboratory manuals Proteins analysis Laboratory Manuals Proteins isolation & purification Laboratory Manuals Methode (DE-588)4038971-6 gnd Proteine (DE-588)4076388-2 gnd Biochemie (DE-588)4006777-4 gnd |
| topic_facet | Eiwitsynthese Eiwitten Proteínas (análise;isolamento e purificação) Protéines - Purification - Manuels de laboratoire Scheidingstechnieken Spectrometrie Proteins Purification Laboratory manuals Proteins analysis Laboratory Manuals Proteins isolation & purification Laboratory Manuals Methode Proteine Biochemie |
| url | http://www.loc.gov/catdir/description/wiley0310/2003012325.html http://www.loc.gov/catdir/toc/wiley031/2003012325.html http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=012818185&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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