Stains Well for Cytoplasmic TCF1

Charite Strasse 1
Deutsches Rheuma-Forschungszentrum
Research Scientist

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Company:

Biolegend

Product Name:

Alexa Fluor® 647 anti-TCF1 (TCF7) Antibody

Catalog Number:

655204

Clone Number:

7F11A10

Info: Supplier Page View Product Specs

Browse Similar Products: TCF1 Antibodies

Our research aims at understanding how some CD4 T cells become autoreactive in melanoma patients treated with check point inhibitors. CD4 T cells are usually identified as CD3+ CD4+ cells. In this experiment, we used anti-TCF-1 Al647 antibody to identify TCF1+ CD4 T cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

PMA+ionomycin+Calcium-activated healthy donor's PBMCs

Primary Incubation

Surface staining with anti-CD3, anti-CD4 etc... (20' at 4°C, in the dark).

Blocking Agent

None

Secondary Incubation

Staining with L/D dye (20' at 4°C, in the dark).

Tertiary Incubation

Cells were fixed and permeabilized with a commercial kit to counterstain for cytoplasmic proteins, including cytokines. The anti-TCF1 Al647 was added on the cells at 1/50 in perm/wash buffer and incubated for 30' at 4°C, in the dark.

Detection

Red laser on Symphony cytometer

Results Summary

Cytoplasmic TCF-1 can be seen on CD4+ CD3+ T cells even without nuclear permeabilization.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

The antibody is specific.

The Bad

The antibody gives good signal, but a FMO control is important to set the positive threshold.

The Bottom Line

Can be used

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