Laboratory methods in enzymology: Protein: C
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Amsterdam [u.a.]
Elsevier, Acad. Press
2014
|
| Ausgabe: | 1. ed. |
| Schriftenreihe: | Methods in enzymology
541 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XVII, 272 S. Ill., graph. Darst. |
| ISBN: | 9780124201194 |
Internformat
MARC
| LEADER | 00000nam a2200000 cc4500 | ||
|---|---|---|---|
| 001 | BV041996890 | ||
| 003 | DE-604 | ||
| 005 | 00000000000000.0 | ||
| 007 | t | ||
| 008 | 140728s2014 ad|| |||| 00||| eng d | ||
| 020 | |a 9780124201194 |9 978-0-12-420119-4 | ||
| 035 | |a (OCoLC)885227750 | ||
| 035 | |a (DE-599)BVBBV041996890 | ||
| 040 | |a DE-604 |b ger |e rakwb | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-355 |a DE-19 | ||
| 245 | 1 | 0 | |a Laboratory methods in enzymology: Protein |n C |
| 250 | |a 1. ed. | ||
| 264 | 1 | |a Amsterdam [u.a.] |b Elsevier, Acad. Press |c 2014 | |
| 300 | |a XVII, 272 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 1 | |a Methods in enzymology |v 541 | |
| 490 | 0 | |a Methods in enzymology |v ... | |
| 650 | 0 | 7 | |a Enzymologie |0 (DE-588)4152482-2 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Labortechnik |0 (DE-588)4123602-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Proteine |0 (DE-588)4076388-2 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4143413-4 |a Aufsatzsammlung |2 gnd-content | |
| 689 | 0 | 0 | |a Enzymologie |0 (DE-588)4152482-2 |D s |
| 689 | 0 | 1 | |a Proteine |0 (DE-588)4076388-2 |D s |
| 689 | 0 | 2 | |a Labortechnik |0 (DE-588)4123602-6 |D s |
| 689 | 0 | |5 DE-604 | |
| 700 | 1 | |a Lorsch, Jon |e Sonstige |4 oth | |
| 773 | 0 | 8 | |w (DE-604)BV041718771 |g 3 |
| 830 | 0 | |a Methods in enzymology |v 541 |w (DE-604)BV000000938 |9 541 | |
| 856 | 4 | 2 | |m Digitalisierung UB Regensburg - ADAM Catalogue Enrichment |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027439011&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-027439011 | ||
Datensatz im Suchindex
| _version_ | 1804152404773437440 |
|---|---|
| adam_text | CONTENTS
Contributors
xiii
Preface
xvii
Section I
Protein Protocols/Protein Precipitation
1.
TCA Precipitation
3
Laura Koontz
Ί.
Theory
4
2.
Equipment
4
3.
Materials
4
4.
Protocol
6
5.
Step 1A Trichloroacetic Acid Precipitation
6
6.
Step 1B Deoxycholate-Trichloroacetic Acid Precipitation
7
References
10
Section II
Protein Protocols/Protein Pull-Down Methods
2.
Coimmunoprecipitation of Proteins from Yeast
13
Erica Gerace and Danesh Moazed
1.
Theory
14
2.
Equipment
15
3.
Materials
15
4.
Protocol
17
5.
Step
1
Preparation of Whole Cell Lysates
18
6.
Step
2
Normalization of Cell Lysates
20
7.
Step
3
Coimmunoprecipitation
20
8.
Step
4
Wash and Elute the Immunoprecipitates
23
9.
Step
5
Analysis of Immunoprecipitations
24
References
26
3.
Coupling Antibody to Cyanogen Bromide-Activated Sepharose
27
Jennifer M. Kavran and Daniel J. Leahy
1.
Theory
28
2.
Equipment
28
vi
Contents
3.
Materials
28
4.
Protocol
30
5.
Step
1
Preparation of Antibody and Resin
30
6.
Step
2
Coupling the Antibody to the Resin
32
7.
Step
3
Quench the Reaction
32
8.
Step
4
Wash the Resin
33
References
34
4.
Analysis of Protein-Protein Interactions by Coimmunoprecipitation
35
Peter van
der Geer
1.
Theory
36
2.
Equipment
38
3.
Materials
39
4.
Protocol
42
5.
Stepi
Isolation of the Protein of Interest by Immunoprecipitation
42
6.
Step
2
Detection of the Binding Partner by Immunoblotting
44
References
46
Section III
Protein Protocols/Protein Purification
5.
Use and Application of
Hydrophobie
Interaction Chromatography
for Protein Purification
51
Justin T. McCue
1.
Theory
52
2.
Equipment
56
3.
Materials
56
4.
Protocol
58
5.
Step
1
Column Equilibration
59
6.
Step
2
Column Loading
60
7.
Step
3
Product Elution
61
8.
Step
4
Adsorbent Regeneration and Sanitization
63
References
64
Source References
65
6.
Hydroxyapatite Chromatography: Purification Strategies for
Recombinant
Proteins
67
Larry J. Cummings
1.
Theory
68
2.
Equipment
69
Contents_____________________________________________________________________v[i
3.
Materiais
69
4.
Protocol
72
5.
Step
1
Purification Protocol Screening by Linear Salt Gradient
75
6.
Step
2
Purification Protocol Using a Step Gradient
77
7.
Step
3
Purification Protocol Using a Step Gradient and Simplified Analytics
79
8.
Step
4
Offline
pH
Measurement and Caicium ion Analysis
80
9.
Step
5
SEC Profile for the Collected mAb Fraction and
Regeneration Fraction
81
References
83
7.
Salting out of Proteins Using Ammonium
Sulfate
Precipitation
85
Krisna
С.
Duong-Ly and Sandra
В.
Gabelli
1.
Theory
86
2.
Equipment
87
3.
Materials
88
4.
Protocol
88
5.
Step
1
Removal of Proteins Marginally Soluble in (NH4)2SO4
89
6.
Step
2
Precipitation of the Protein of Interest
89
References
94
8.
Using Ion Exchange Chromatography to Purify a Recombinantly
Expressed Protein
95
Krisna
С.
Duong-Ly and Sandra
В.
Gabelli
1.
Theory
96
2.
Equipment
98
3.
Materials
98
4.
Protocol
99
5.
Step
1
Equilibration of the Column
99
6.
Step
2
Binding of the Protein Sample
101
7.
Step
3
Removal of Unbound Proteins
101
8.
Step
4
Elution of the Bound Protein
102
References
103
9.
Gel Filtration Chromatography (Size Exclusion Chromatography)
of Proteins
105
Krisna
С.
Duong-Ly and Sandra
В.
Gabelli
Ί
.
Theory
106
2.
Equipment
108
3.
Materials
108
Vili
Contents
4.
Protocol
109
5.
Step
1
Standardization of the Gel Filtration Column
ì IO
6.
Step
2
Determination of the Sizes of Protein Species in a Sample
112
References
114
Section IV
Protein Protocols/Purification of Membrane Proteins
10.
Expression and Purification of Membrane Proteins
117
Jan Kubíček,
Helena Block, Barbara Maertens, Anne Spriestersbach
and
Jörg Labahn
1.
Theory
118
2.
Equipment
121
3.
Materials
121
4.
Protocol
124
5.
Step
Ί
Transformation off.
coli
126
6.
Step
2
Cultivation of
E. coli
-
Screening for the Optimal
Expression Conditions
126
7.
Step
3
Scale-Up Expression of a Membrane Protein Using
the Optimal Expression Conditions
130
8.
Step
4
Screening Detergents to Determine Optimal Solubilization
of Membrane Protein
131
9.
Step
5
Scale-Up the Solubilization of the Membrane Protein
134
10.
Step
6
Purification of Membrane Proteins Using Ni-NTA Superflow
136
References
140
11.
Explanatory Chapter: Choosing the Right Detergent
141
Dirk Linke
1.
Theory
142
2.
Equipment
142
3.
Materials
142
4.
Protocol
144
5.
Using Detergents with Polyacrylamide Gel Electrophoresis
145
6.
Using Detergents in Chromatograpy
146
7.
Using Detergents with Optical Spectroscopy Techniques
147
8.
Using Detergents with Mass Spectrometry Techniques
147
9.
Using Detergents with Nuclear Magnetic Resonance (NMR)
148
Contents______________________________________________________________________ix
10.
Using Detergents in Protein Crystallization
148
References
148
Section V
Protein Protocols/SDS PAGE
12.
One-dimensional SDS-Polyacrylamide Gel Eiectrophoresis
(lDSDS-PAGE)
151
Julie
L
Brunelle
and Rachel Green
1.
Theory
152
2.
Equipment
152
3.
Materials
153
4.
Protocol
154
5.
Step
1
Casting an SDS-PAGE Gel: Resolving Gel
154
6.
Step
2
Casting an SDS-PAGE Gel: Stacking Gel
155
7.
Step
3
Running an SDS-PAGE
Gei
157
References
159
13.
Coomassie Blue Staining
161
Julie L.
Brunelle
and Rachel Green
1.
Theory
162
2.
Equipment
162
3.
Materials
162
4.
Protocol
163
5.
Step
1
Stain a Gel Using Coomassie Blue
163
6.
Step
2
Destain the Gel to Reduce Background Staining
165
Source References
167
14.
Silver Staining of SDS-polyacrylamide Gel
169
Jennifer M. Kavran and Daniel J. Leahy
170
170
170
172
172
173
175
References
176
1.
Theory
2.
Equipment
3.
Materials
4.
Protocol
5.
Step
1
Fix the Gel
6.
Step
2
Stain the Gel
7.
Step
3
Preserve the Gel
χ
Contents
Section
VI
Protein Protocols/Standard in
vitro
Assays
for Protein-Nucleic Acid Interactions
15.
Standard In Vitro Assays for Protein-Nucleic Acid
Interactions
-
Gel Shift Assays for
RNA
and
DNA
Binding
179
Sarah F. Mitchell and Jon R. Lorsch
1.
Theory
180
2.
Equipment
Τ
84
3.
Materials
184
4.
Protocol
186
5.
Step
1
Rad/olabeling the Nucleic Acid Probe
187
6.
Step
2
Bind Protein and Nucleic Acid
188
7.
Step
3
Preparation of Polyacrylamide Gel
190
8.
Step
4
Loading and Running Gel
191
9.
Step
5
Analysis of Gel
192
References
195
Source References
196
16.
Protein Filter Binding
197
Sarah Kolitz and Jon R. Lorsch
1.
Theory
198
2.
Equipment
198
3.
Materials
198
4.
Protocol
199
5.
Step
1
Assemble Binding Reactions
201
6.
Step
2
Quantify Binding
202
7.
Step
3
Process Binding Data
204
References
205
Source References
205
Section
VII
Protein Protocols/Troubleshooting Protein Expression
17.
Explanatory Chapter: Troubleshooting
Recombinant
Protein Expression: General
209
Krisna
С.
Duong-Ly and Sandra
В.
Gabelli
1.
Theory
210
2.
Equipment
213
3.
Materiais
213
Contents
X¡
4.
Protocol
214
5.
Step
1
Monitoring
£
coli
cell Growth Before Induction
214
6.
Step
2
Induction of Expression
216
7.
Step
3
Monitoring
£
¿го//
Cell Growth After Induction
217
8.
Step
4
Measurement of Protein Production
218
9.
Step
5
Further Troubleshooting in
E. coli
219
TO. Step
б
Eukaryotic Expression Systems
223
References
227
Source References
229
18.
Explanatory Chapter: Troubleshooting Protein Expression:
What to do When the Protein is not Soluble
231
Krisna
С.
Duong-Ly and Sandra
В.
Gabelli
1.
Theory
232
2.
Equipment
232
3.
Materials
233
4.
Protocol
234
5.
Step
1
Centrifuge the Culture
235
6.
Step
2
Lyse
the Cells
235
7.
Step
3
Remove the Cell Debris
237
8.
Step
4
Analyze Protein Expression by SDS-PAGE
237
9.
Step
5
Troubleshooting the Lack of Soluble Protein Expressed
239
References
244
Section
VIII
Protein Protocols/Western Blotting
19.
Western Blotting using Chemiluminescent Substrates
251
Alice Alegria-Schaffer
1.
Theory
252
2.
Equipment
252
3.
Materials
252
4.
Protocol
253
5.
Step
1:
Protein Transfer to a Membrane
254
6.
Step
2:
Western Blot Detection using a Chemiluminescent Substrate
256
References
258
Source References
259
Author Index
261
Subject Index
267
|
| any_adam_object | 1 |
| building | Verbundindex |
| bvnumber | BV041996890 |
| ctrlnum | (OCoLC)885227750 (DE-599)BVBBV041996890 |
| edition | 1. ed. |
| format | Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01651nam a2200421 cc4500</leader><controlfield tag="001">BV041996890</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">00000000000000.0</controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">140728s2014 ad|| |||| 00||| eng d</controlfield><datafield tag="020" ind1=" " ind2=" "><subfield code="a">9780124201194</subfield><subfield code="9">978-0-12-420119-4</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)885227750</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)BVBBV041996890</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-355</subfield><subfield code="a">DE-19</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Laboratory methods in enzymology: Protein</subfield><subfield code="n">C</subfield></datafield><datafield tag="250" ind1=" " ind2=" "><subfield code="a">1. ed.</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Amsterdam [u.a.]</subfield><subfield code="b">Elsevier, Acad. Press</subfield><subfield code="c">2014</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">XVII, 272 S.</subfield><subfield code="b">Ill., graph. Darst.</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="490" ind1="1" ind2=" "><subfield code="a">Methods in enzymology</subfield><subfield code="v">541</subfield></datafield><datafield tag="490" ind1="0" ind2=" "><subfield code="a">Methods in enzymology</subfield><subfield code="v">...</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Enzymologie</subfield><subfield code="0">(DE-588)4152482-2</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Labortechnik</subfield><subfield code="0">(DE-588)4123602-6</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Proteine</subfield><subfield code="0">(DE-588)4076388-2</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4143413-4</subfield><subfield code="a">Aufsatzsammlung</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="689" ind1="0" ind2="0"><subfield code="a">Enzymologie</subfield><subfield code="0">(DE-588)4152482-2</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2="1"><subfield code="a">Proteine</subfield><subfield code="0">(DE-588)4076388-2</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2="2"><subfield code="a">Labortechnik</subfield><subfield code="0">(DE-588)4123602-6</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2=" "><subfield code="5">DE-604</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Lorsch, Jon</subfield><subfield code="e">Sonstige</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="w">(DE-604)BV041718771</subfield><subfield code="g">3</subfield></datafield><datafield tag="830" ind1=" " ind2="0"><subfield code="a">Methods in enzymology</subfield><subfield code="v">541</subfield><subfield code="w">(DE-604)BV000000938</subfield><subfield code="9">541</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">Digitalisierung UB Regensburg - ADAM Catalogue Enrichment</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027439011&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-027439011</subfield></datafield></record></collection> |
| genre | (DE-588)4143413-4 Aufsatzsammlung gnd-content |
| genre_facet | Aufsatzsammlung |
| id | DE-604.BV041996890 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T01:10:11Z |
| institution | BVB |
| isbn | 9780124201194 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-027439011 |
| oclc_num | 885227750 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-19 DE-BY-UBM |
| owner_facet | DE-355 DE-BY-UBR DE-19 DE-BY-UBM |
| physical | XVII, 272 S. Ill., graph. Darst. |
| publishDate | 2014 |
| publishDateSearch | 2014 |
| publishDateSort | 2014 |
| publisher | Elsevier, Acad. Press |
| record_format | marc |
| series | Methods in enzymology |
| series2 | Methods in enzymology |
| spelling | Laboratory methods in enzymology: Protein C 1. ed. Amsterdam [u.a.] Elsevier, Acad. Press 2014 XVII, 272 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Methods in enzymology 541 Methods in enzymology ... Enzymologie (DE-588)4152482-2 gnd rswk-swf Labortechnik (DE-588)4123602-6 gnd rswk-swf Proteine (DE-588)4076388-2 gnd rswk-swf (DE-588)4143413-4 Aufsatzsammlung gnd-content Enzymologie (DE-588)4152482-2 s Proteine (DE-588)4076388-2 s Labortechnik (DE-588)4123602-6 s DE-604 Lorsch, Jon Sonstige oth (DE-604)BV041718771 3 Methods in enzymology 541 (DE-604)BV000000938 541 Digitalisierung UB Regensburg - ADAM Catalogue Enrichment application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027439011&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Laboratory methods in enzymology: Protein Methods in enzymology Enzymologie (DE-588)4152482-2 gnd Labortechnik (DE-588)4123602-6 gnd Proteine (DE-588)4076388-2 gnd |
| subject_GND | (DE-588)4152482-2 (DE-588)4123602-6 (DE-588)4076388-2 (DE-588)4143413-4 |
| title | Laboratory methods in enzymology: Protein |
| title_auth | Laboratory methods in enzymology: Protein |
| title_exact_search | Laboratory methods in enzymology: Protein |
| title_full | Laboratory methods in enzymology: Protein C |
| title_fullStr | Laboratory methods in enzymology: Protein C |
| title_full_unstemmed | Laboratory methods in enzymology: Protein C |
| title_short | Laboratory methods in enzymology: Protein |
| title_sort | laboratory methods in enzymology protein |
| topic | Enzymologie (DE-588)4152482-2 gnd Labortechnik (DE-588)4123602-6 gnd Proteine (DE-588)4076388-2 gnd |
| topic_facet | Enzymologie Labortechnik Proteine Aufsatzsammlung |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027439011&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV041718771 (DE-604)BV000000938 |
| work_keys_str_mv | AT lorschjon laboratorymethodsinenzymologyproteinc |