Extremely Strong Antibody Against GST

Cell, Molecular and Cancer Biology
University of Massachusetts Medical School
Post Doctoral Research Associate

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Santa Cruz Biotechnology

Product Name:

GST Antibody (B-14): sc-138

Catalog Number:

sc-138

Info: View Product View Product Specs

Browse Similar Products: GST Antibodies

The aim of this experiment was to study the function of a variety of yeast G1 cyclins by expressing them in budding yeast Saccharomyces cerevisiae as C-terminal GST-tagged proteins and using a common substrate. Here the exression of GST tagged cyclins are detected successfully

Experimental Design and Results Summary

Applications

Western Blot

Sample

Cell Lysates from budding yeast

Primary Incubation

1:1000 Dilution in 5% Milk (TTBS), overnight incubation in cold room

Blocking Agent

5% non-fat milk in TBS-T

Secondary Incubation

Anti-Mouse-HRP, 1:3000, 1hour incubation at Room Temperature

Tertiary Incubation

None

Detection

Bio-Rad Clarity HRP substrate

Results Summary

Protein extract from cells was prepared in TCA lysis buffer supplied with PMSF, protease inhibitor mixture, and sodium orthovanadate. Lysates were quantified using a BCA assay kit (Thermos). 20 micrograms of proteins were subjected to SDS-PAGE and transferred onto PVDF membranes and blocked with 5% non-fat milk in TBS-T (20 mMTris, 500 mMNaCl, and 0.1% Tween 20) at room temperature for 1 h with rocking. The membranes were probed with primary Antibody (1/1000 dilution) overnight at 4°C. After washing with TBS-T, the membranes were incubated with HRP-conjugated secondary antibody in 5% non-fat milk/TBS-T at room temperature for 1 h. The protein–Antibody complex was detected by Bio-Rad Clarity ECL substrate. The antibody detected GST-tagged cyclins robustly.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Works great for GST

The Bad

None

The Bottom Line

I would use this antibody for detection of GST in any cells.

Share your experience with other scientists. Write a Review! »

Join the discussion