Antibody to Detect Bcl-6

October 06, 2017

Molecular and Cell Biology
University of California, Merced
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Invitrogen Antibodies

Product Name:

BCL6 Monoclonal Antibody (BCL-DWN), APC, eBioscience™

Catalog Number:

17-5453-80

We use Bcl-6 to investigate CD4 T follicular helper cells. Invitrogen suggests to use this antibody at a fairly dilute concentration so it can carry through a large number of experiments. However, it can be hard to detect if it is not highly upregulated in the population of interest. There may be better antibodies for detection but this works well for the levels of Bcl-6 that we want to detect in T follicular helper cells

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine Lymphocytes

Primary Incubation

Any surface markers of interest 30 min at 4 degrees

Blocking Agent

FC Block

Secondary Incubation

FoxP3 Fixation 45 min at room temperature

Tertiary Incubation

1:1000 dilution of Bcl-6 in Perm Buffer 45 min at room temperature

Detection

BD LSR II

Results Summary

Bcl-6 can be detected by a 1:1000 dilution of Bcl-6 when analyzing T follicular helper cells. It is most noticeable as a shift as opposed to a distinct population.

DOI or PMID #

N/A

Additional Notes

The FoxP3 fixation buffer works best for my detection of Bcl-6

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Summary

The Good

It works well at a reasonable dilution.

The Bad

Results appear as more than a shift as opposed to distinct populations.

The Bottom Line

It will work well for a Bcl-6+ population but it may need to be further optimized to distinguished cells with a faint signal.

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