Quality GAPDH Antibody

December 21, 2015

Research
University of North Carolina at Chapel Hill
Laboratory Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

GeneTex

Product Name:

GAPDH antibody [GT239]

Catalog Number:

GTX627408

Our lab aims to elucidate the role of kinases in several cancers, and we are often performing Western Blots to determine kinase protein and activity levels in various cell lines. GAPDH is one of our favorite loading controls for which to blot because of its convenient 36 kDa size and extremely low background on film. We chose this antibody because of a recommendation from another lab, and have found it to be quite useful ever since.

Experimental Design and Results Summary

Applications

Western Blot

Sample

786-0, RPTEC, and RCC4 Cell Lysates

Primary Incubation

1:5000 Dilution Overnight in 5% BSA in 1X TBS-T at 4C

Blocking Agent

5% Milk in 1X TBS-T

Secondary Incubation

1:5000 dilution of HRP-conjugated Rabbit secondary in 5% Milk in TBS-T for 1 hour at RT

Tertiary Incubation

NA

Detection

ECL --> Autoradiography Film via HRP-Conjugate Secondary Antibody

Results Summary

Our lab has been able to demonstrate consistently that our Western Blots have been evenly loaded using this antibody to GAPDH. Using autoradiography film and a low-strength mixture of ECL, there was very little background and a strong signal. Around 10-15 seconds was sufficient to produce bands on film that were strong but not overpowering.

Additional Notes

Base ECL strength is plenty for this antibody to produce bands on film.

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Summary

The Good

Strong signal, low background

The Bad

Expensive

The Bottom Line

Having a reliable, efficient loading control is essential for a well-developed Western Blot. Our lab makes use of several, but this antibody to GAPDH is likely my favorite because of how clean it is, and how little needs to be used. I highly recommend this GAPDH antibody to anyone else needing an accurate loading control for their Western Blots.

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