Proteomics & Protien Expression

Preparing GST Fusion Proteins

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Selected Protocol from "Protein-Protein Interactions", 2nd Edition, E. Golemis, P. Adam.

Preparation of Glutathione-S-Transferase (GST) Fusion Proteins

Margret B. Einarson and Elena N. Pugacheva
Foxx Chase Cancer Center, Philadelphia, PA 19111

Jason R. Orlinck
Brigham & Women’s Hospital, Boston, MA 02115


ABSTRACT
Glutathione-S-Transferase (GST) fusion proteins have a wide range of applications. This protocol is designed for IPTG-inducible bacterial expression vectors.  

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Products Available for this Protocol
Protocol Material Description Product #  Product Name Add to Cart
Reduced glutathione, 20 mM prepared in 50 mM Tris-HCl (pH 8.0) G4251 L-Glutathione reduced
Isopropyl-β-D-thio-galactoside (IPTG) I5502 Isopropyl β-D-1-thiogalactopyranoside
LB broth, containing appropriate antibiotic selection L2542 LB Broth
PBS P5368 Phosphate buffered saline
1% Triton X-100 X-100 Triton® X-100
Aprotinin A1153 Aprotinin from bovine lung
Leupeptin L9783 Leupeptin hydrochloride
Phenylmethylsulfonyl fluoride [PMSF] P7626 Phenylmethanesulfonyl fluoride
150 mM NaCl S3014 Sodium chloride
20 mM sodium phosphate (pH 7.4) S3264 Sodium phosphate dibasic
Glutathione-Sepharose beads G4510 Glutathione−Agarose
Coomassie blue (see Sambrook and Russell 2001) B5133 Brilliant Blue G
100 mM Tris-HCl (pH 6.8) T3253 Trizma® hydrochloride
4%(w/v) SDS (electrophoresis grade) L4390 Sodium dodecyl sulfate
0.2% (w/v) bromophenol blue B0126 Bromophenol Blue
20% (v/v) glycerol G9012 Glycerol
200 mM dithiothreitol (DTT) D9779 DL-Dithiothreitol
β-mercaptoethanol M3148 2-Mercaptoethanol

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