Phosphorylated eIF2a to Detect ER Stress

Food Science and Human Nutrition
Colorado State University
Graduate Student

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Company:

Abcam

Product Name:

Anti-EIF2S1

Catalog Number:

ab32157

eIF2a is a well known substrate of at least 4 kinases, the most well known being PERK. When, phosphorylated, it has been demonstrated to reduce global protein translation, a critical step in reversing ER stress. Our goal was to use this antibody for western blot to determine the amount of phosphorylated eIF2a in a variety of samples treated with various ER stress inducers.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Total cell lysates of H4IIE rat hepatoma cells

Primary Incubation

1:500 antibody in 1:3 Odyssey Block: TBST overnight, at 4 degrees C.

Blocking Agent

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

Secondary Incubation

1:15000 IRDye800CW in 1:3 Odyssey Block: TBST

Tertiary Incubation

N/A

Detection

Licor Odyssey Clx

Results Summary

This antibody works well and shows up at around 36kDa. There is an obvious difference between the treatments. These were compared to total eIF2a.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Minimal background, and strong antibody signal using abcam's dilution recommendation

The Bad

N/A.

The Bottom Line

Order this antibody if you want clear, distinct p-eIF2a results with minimal background

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