Western Blot of HeLa Whole-Cell Lysates Using GAPDH Antibody

February 21, 2019

Laboratory of t-RNA Transcription
Centre for Cellular and Molecular Biology
Research Associate

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

GAPDH (14C10) Rabbit monoclonal antibody

Catalog Number:

2118

Our aim was to examine the levels of phospho-AKT in scramble siRNA and target siRNA transfected HeLa cells. Whole-cell lysates were run on SDS-PAGE and transferred onto PVDF membrane. GAPDH antibody was used as a loading control.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa whole-cell lysate

Primary Incubation

1:10000, overnight at 4°C

Blocking Agent

5% skimmed milk, 1 hr at room temperature

Secondary Incubation

1:10000, anti-rabbit HRP conjugated antibody, 1 hr at room temperature

Tertiary Incubation

N/A

Detection

ECL reagent, Chemi doc

Results Summary

We observed a single specific band of GAPDH around 35 kDa in both control and target siRNA treated HeLa cells without any non-specific bands on western blot.

DOI or PMID #

29143563

Additional Notes

N/A

Image Gallery

Summary

The Good

Excellent antibody for performing western blots

The Bad

N/A

The Bottom Line

An excellent antibody which gives a specific band at 37 kDa. Antibody dilution can be used multiple times.

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