Ga s Antibody for Western Blotting

Biochemistry
Stanford University
Senior Research Scientist

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Company:

Santa Cruz Biotechnology

Product Name:

Gα s Antibody

Catalog Number:

sc-823

The Gα s subunit of the heterotrimeric G proteins stimulates adenylate cyclases to generate cAMP, an important second messenger molecule that is necessary for PKA activation. The goal of this experiment is to test if NIH3T3 cells express Gα s. If it is expressed, I would then go on to determine the localization of this protein. The reason for selecting this product from Santa Cruz is that it was referenced in few publications.

Experimental Design and Results Summary

Applications

Western Blot

Sample

NIH3T3 cells_50 µg of whole cell extract

Primary Incubation

1:200 in blocking agent for 16 hr at 4 deg

Blocking Agent

5% milk powder in 1x TBS-T (0.1% Tween-20)

Secondary Incubation

Donkey anti-Rabbit IgG HRP

Tertiary Incubation

NA

Detection

ECL pico for 5 min

Results Summary

This antibody detects a band at ~46 kDa (expected size) but also detects few other bands at 50 and 55 kDa. The signal is poor and one needs ECP pico or ECL femto. I had tried changing milk powder with BSA in blocking buffer but did not improve the result. I had also tried primary antibody incubation for 1 hr at room temperature but even that did not improve the results.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

OK antibody

The Bad

Poor signal with many background bands

The Bottom Line

Not the best in the market

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