Alpha Tubulin Santa Cruz Biotech

March 08, 2016

Food Science and Human Nutrition
Colorado State University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Santa Cruz Biotechnology

Product Name:

Alpha Tubulin (H-300)

Catalog Number:

sc-5546

We wanted to use this antibody for western blot as well as fluorescent microscopy to assess the amount of total vs acetylated tubulin, each important cytoskeletal components of a cell.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Total cell lysates of H4IIE rat hepatoma cell line

Primary Incubation

1:500 in 1:3 Odyssey Block: TBS over night at 4 degrees C.

Blocking Agent

1:3 Odyssey Block:TBS for 1 hour at room temperature

Secondary Incubation

1:3 Odyssey Block: TBST (0.1% Tween, 0.02% SDS) for 1 hour at room temperature

Tertiary Incubation

NA

Detection

Odyssey Infrared Imager

Results Summary

This antibody is bad. The signal was inconsistent and weak. Oddly the acetylated tubulin that was multiplexed with this antibody showed a stronger, better, more consistent band which should not have been the case.

Additional Notes

None

Related Categories

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Summary

The Good

It worked...

The Bad

The signal was so weak I could not trust this antibody. It was very inconsistent, making this antibody useless as a measurable control.

The Bottom Line

Don't use this antibody. Santa Cruz has another tubulin antibody that is much more reliable (sc-5286)

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