alpha-Tubulin Antibody for Western Blot

February 23, 2018

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 Antibody, Mouse monoclonal

Catalog Number:

SC5286

This antibody was selected to function as a loading control to normalize protein expression to. The specific application I use it for is multiplexing with rabbit antibodies in order to normalize protein expression to alpha-tubulin on one membrane.

Experimental Design and Results Summary

Applications

Western Blot

Sample

H4IIE Whole cell lysates

Primary Incubation

1:1000, 4C, overnight with gentle shaking in 1:3 Odyssey Blocking buffer (Licor):TBST (0.1% tween20)

Blocking Agent

1:3 Odyssey Blocking buffer:TBS

Secondary Incubation

1:15,000 goat-anti-mouse (Licor IRDye800CW) at room temperature with gentle shaking in 1:3 Odyssey Blocking buffer (Licor):TBST (0.1% tween20, 0.02% SDS)

Tertiary Incubation

N/A

Detection

Odyssey Clx

Results Summary

This antibody produces a strong band around 55 kda with minimal non-specific binding. It works well, and is consistent.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Good reliable antibody to use a a loading control for western blot

The Bad

Detects one random band around 90 kDa

The Bottom Line

Great antibody to use as a loading control. Produces consistent results with minimal background signal.

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