CHOP (DDIT3) Antibody

February 09, 2018

Food Science and Human Nutrition
Colorado State University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

anti-DDIT3 (CHOP) antibody [9C8]

Catalog Number:

ab11419

This antibody was used to detect the presence of endoplasmic reticulum stress / unfolded protein response activation, specifically the IRE1 arm. This antibody used to validate gene expression of CHOP. It was used with a positive control for UPR activation, thapsigargin, and produced good, reliable results.

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 works well, it requires a good bit of protein (loaded 30ug). It works well for a marker of ER stress/UPR activation. Detection is around 31kDa.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Great marker for ER stress

The Bad

Need a good bit of ER stress going on to detect it

The Bottom Line

Great antibody to detect for presence of ER stress/UPR induction. If you treat with something other than a chemical ER stress inducer, you will probably need to load a fair amount of protein.

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