GST Purification Kit

Stanford University
Biochemistry
Senior Research Scientist

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Company:

Bio-Rad

Product Name:

GST Purification Kit

Catalog Number:

6200244

Rab GTPases are small GTPases of the Ras superfamily that regulate multiple processes in the secretory pathway. They cycle between GTP bound (active) and GDP bound (inactive) states. GEF proteins regulate GDP-GTP exchange and GAP proteins regulate GTP hydrolysis. Ric1-Rgp1 complex is a GEF complex that regulates Rab6.In the present experiment, a fragment of Ric1 protein was purified for binding assays. This fragment is a GST tagged protein.

Experimental Design and Results Summary

Application

Recombinant protein purification

Starting Material

Bacterially expressed protein

Protocol Overview

GST-tagged Ric1 fragment was expressed in E coli BL21 cells by induction with 0.1 mM IPTG for 4 hr at room temp. Cells were harvested and lysed in lysis buffer (50 mM Tris-HCl, pH 8.0, 200 mM NaCl, 5 mM MgCl2, 0.1 mM dithiothreitol (DTT), 1 mM PMSF). Cellslysis was done by a single pass at 20,000 p.s.i. through an EmulsiFlex-C5 (Avestin). Samples were spun at 50,000xg for 30 min at 4 deg. Clarified samples were then used for protein purification according to the manufacturer’s instructions.

Tips

Follow the instructions from the manufacturer and it works well.

Results Summary

GST-tagged Ric1 fragment was successfully purified without any major contaminants.

Additional Notes

None

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Summary

The Good

Easy to use.

The Bad

None

The Bottom Line

One of the best GST protein purification systems in the market

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