BioLegend BV421 Anti-human CD45RA Antibody Works Well

July 18, 2019

JvGCRC
Nottingham Trent University
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Brilliant Violet 421™ anti-human CD45RA Antibody

Catalog Number:

304129

Clone Number:

HI100

This antibody has been used to optimise flow cytometry-based immunophenotyping of whole-blood samples. The results have not yet been published. Multi-colour antibody panels were designed to identify a number of lymphocyte populations and tested to ensure appropriate compensation was possible. CD45RA was used in a ten-colour panel to help distinguish memory, effector and naive T cells. It is easy to use and gives clear staining.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human whole blood

Primary Incubation

3µL BV421-CD45RA for 30 minutes at 4°C

Blocking Agent

FcR blocking reagent, human (Miltenyi Biotec, cat# 130-059-901)

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

Beckman Coulter Gallios, Kaluza Software

Results Summary

Clear staining that, in combination with CD62L, allowed the distinction of memory, effector and naive T cell populations. Titration optimisations showed we could reduce the volume of antibody used with no detriment to the results. Prior to analysing samples, erythrocytes were lysed using 1.5 mL 1xPharmLyse (BD, cat#555899) at room temperature for 30 minutes. This antibody fit well into a nine-colour panel with no compensation issues.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Useful for identifying memory, effector and naive T cell populations in multi-colour panels, and can be titrated from the manufacturers suggested volume.

The Bad

The antibody has only been tested for flow cytometry.

The Bottom Line

Easy-to-use pack that provides clear staining.

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