CHOP Protein Detection Via Western Blot

April 12, 2016

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 antibody

Catalog Number:

ab11419

CHOP is a downstream marker of the unfolded protein response (UPR). Increased expression of CHOP indicates ER stress and activation of the UPR. Therefore, being able to quantitatively measure CHOP levels in total cell lysates is critical to determine the level of ER stress due to a variety of treatment conditions.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Total cell lysates of rat hepatoma cells

Primary Incubation

1:1000 antibody in 0.1% tween and 1:3 odyssey block: 1X TBS at 4 degrees C overnight.

Blocking Agent

1:3 odyssey block:1X TBS for 1 hour room temperature

Secondary Incubation

1:15,000 IRDye 680RD from Li-Cor Biosciences in 0.1% tween, 0.02% SDS, in 1:3 odyssey block: 1X TBS for 1 hour room temperature

Tertiary Incubation

NA

Detection

Odyssey Infrared Imaging System

Results Summary

This is the first (of many) CHOP antibody we have tested that worked! There is a clear difference in CHOP levels in each treatment group. This antibody is definitely what we wanted.

Additional Notes

None

Related Categories

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Summary

The Good

Minimal background and minimal non-specific binding with the recommended dilution

The Bad

EXPENSIVE! Signal could be stronger. Would decrease dilution to 1:500.

The Bottom Line

Abcam CHOP antibody works! Although it is expensive, Abcam has a guarantee that if an antibody doesn't work they will refund your money or exchange the antibody for another and they don't lie! We have spoken to representatives and exchanged a few before and they are easy to work with.

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