EIF2-Alpha Antibody For Western Blot

February 12, 2018

Food Science and Human Nutrition
Colorado State University
Graduate Student

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Company:

Santa Cruz Biotechnology

Product Name:

eIF2-alpha antibody - mouse monoclonal

Catalog Number:

SC133132

This antibody was selected as a control to normalize the ratio of phosphorylated eIF2a to total eIF2-alpha. The context of this application was to measure for activation of the unfolded protein response, specifically activation of the PERK arm of the UPR.

Experimental Design and Results Summary

Applications

Western Blot

Sample

H4IIE Whole cell lysates

Primary Incubation

1:500, 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 at 36 kDa with minimal background via western blot. We have found this antibody to be reliable, and have used it for years in combination with a phospho-eIF2-alpha antibody.

DOI or PMID #

N/A

Additional Notes

Loaded 30ug of protein

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Summary

The Good

Reliable, strong signal, minimal background

The Bad

No major drawbacks

The Bottom Line

Great antibody, well worth the money!

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