Weak CD45RO Staining

Charite Strasse 1
Deutsches Rheuma-Forscungszentrum
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Miltenyi

Product Name:

CD45RO Antibody, anti-human, VioBlue®, REAfinity™

Catalog Number:

130-119-710

Clone Number:

REA611

Info: Supplier Page View Product Specs

Browse Similar Products: CD45RO 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 CD45RO VioBlue antibody to identify memory CD4 and CD8 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-CD45RO VioBlue at 1/100 (see titration), anti-CD45RA, anti-CD3, anti-CD4, anti CD8, 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 cytokines.

Detection

Violet laser on Symphony cytometer

Results Summary

CD45RO can be seen on CD4+ CD3+ T cells but the antibody should be carefully titrated, as it does not work well if too concentrated, and is rapidly lost above 1/100 dilution.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

The antibody is specific, but not ideal on activated cells.

The Bad

This antibody needs careful titration and is not very bright.

The Bottom Line

Can be used.

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