PE/Dazzle 594 Anti-Human HLA-DR Antibody from BioLegend Works Well

July 25, 2019

JvGCRC
Nottingham Trent University
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

PE/Dazzle™ 594 anti-human HLA-DR Antibody

Catalog Number:

307654

Clone Number:

L243

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. HLA-DR was used in a ten-colour panel to assess levels in different lymphocyte populations. It is easy to use and gives clear staining.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human whole blood

Primary Incubation

3µL PE/Dazzle594.HLA-DR 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 was used to confirm HLA-DR expression on a numberof lymphocyte 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 ten-colour panel with no compensation issues.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Clear staining of HLA-DR expression on a variety of cells that is useful for FACS analysis.

The Bad

The antibody has only been tested for flow cytometry.

The Bottom Line

An easy-to-use pack that could be titrated below the manufacturers suggested volume and still provide clear staining.

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