Protocols and Applications Guide (US Letter Size) - Promega

||||||||||| 11Protein Purification and Analysis
CONTENTS
I. Introduction 1
II. Affinity Tags 1
A. Polyhistidine 1
B. Glutathione-S-Transferase 1
III. Purification of Polyhistidine-Tagged Proteins 1
A. Rapid Purification of Polyhistidine-Tagged Proteins Using
Magnetic Resins 1
B. Medium- to Large-Scale Purification of
Polyhistidine-Tagged Proteins In Column or Batch
Formats 5
C. 96-Well Format For Purification of Polyhistidine-Tagged
Proteins 7
IV. Purification of GST-Tagged Proteins 9
A. Rapid Purification of GST-Tagged Proteins Using
Magnetic Resins 9
V. Purification of Biotinylated Proteins 11
A. PinPoint Xa System and SoftLink Resin for
Purification of Biotinylated Protein 11
VI. Protein:Protein Interaction Analysis: In Vivo and In
Vitro Methods 13
A. Mammalian Two-Hybrid Systems 13
B. In Vitro Pull-Down Assays 15
VII. Analysis of DNA:Protein Interactions 19
A. Gel Shift Assays 19
B. Chromatin Immunoprecipitation 20
VIII.Proteomics Approaches for the Analysis of Complex
Mixtures of Proteins 20
A. In-Gel Trypsin Digestion of Proteins 21
B. In-Solution Trypsin Digestion of Proteins 23
C. Affinity Tag In Vitro Pull-Down Assay with Trypsin
Digestion and Protein Analysis 24
IX. SDS-PAGE Analysis 24
X. Composition of Solutions 25
XI. References 26
Protocols & Applications Guide
www.promega.com
rev. 3/09
PROTOCOLS & APPLICATIONS GUIDE