Easy Bcl-2 Detection for Western Blot

December 16, 2022

BlueskyReddit
IKP/RBCT
Robert Bosch Gesellschaft für medizinische Forschung mbH
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Bcl-2 (124) Mouse mAb

Catalog Number:

#15071

Clone Number:

124

Info:Supplier Page View Product Specs

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This mouse monoclonal Bcl-2 antibody from Cell singaling is our standard antibody for the detection of endogenous and overexpressed Bcl-2 in human cell lines via Western Blot. Although durability is not too long and functionality for other methods cannot be confirmed the quality of results is mostly satisfying.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Human cancer cell lines (whole cell lysates) e.g. HEK293, MCF-7

Primary Incubation

1:500 at 4°C overnight

Blocking Agent

5 % BSA in TBST

Secondary Incubation

1:2000, 60 min at RT

Tertiary Incubation

None

Detection

HRP, Chemiluminescence (West Dura Extended Duration Substrate) in Elysia-raytest detector (3 min exposure)

Results Summary

The detection of Bcl-2 using this antibody yields a specific band at the size of approx. 26 kDa for endogeneous Bcl-2. Unspecific bands are rare. We also successfully applied this product for the detection of fluorophore-fused Bcl-2 and several Bcl-2 mutants.

DOI or PMID #

N/A

Additional Notes

We use the antibody in a higher concentration (1:500) than recommended (1:1000) since we found out the results are much better.

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Summary

The Good

No unspecific bands, reproducible results.

The Bad

Durability, higher concentration needed.

The Bottom Line

This antibody works well for Western Blot detection of endogeneous and overexpressed Bcl-2. The functionality for other methods could not be confirmed in our lab.

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