Alpha-Tubulin Antibody for Western Blotting

Biochemistry
Stanford University
Senior Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Sigma-Aldrich

Product Name:

Anti-α-Tubulin antibody, Mouse monoclonal

Catalog Number:

T6199

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I am interested in understanding the mechanism of Hedgehog signaling. My project involves protein biochemistry techniques like protein purification, Western Blotting, enzymatic assays etc. The goal of this experiment is to test if alpha-tubulin antibody can be used as a loading control for Western Blotting experiments. This antibody was selected based on a recommendation from a fellow researcher.

Experimental Design and Results Summary

Applications

Western Blot

Sample

NIH3T3 whole cell extract

Primary Incubation

1:10000 in blocking agent for 16 hr at room temperature

Blocking Agent

5% Milk in 1x TBS-T (0.1% Tween)

Secondary Incubation

Donkey anti-mouse IgG HRP at 1:5000 dilution in blocking agent for 1 hr at room temperature

Tertiary Incubation

NA

Detection

ECL for 2 sec

Results Summary

The antibody picks up a nice band at ~50 kDa (the expected size).

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Specific band detection

The Bad

None

The Bottom Line

An excellent antibody that works at high dilutions and at many incubation times ranging from 30 min-1 hr at room temperature or 8-16 hr at 4 deg.

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