Good APC/Fire™ 750 Anti-Human CD45 Antibody from BioLegend

July 16, 2019

JvGCRC
Nottingham Trent University
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

APC/Fire™ 750 anti-human CD45 Antibody

Catalog Number:

368517

Clone Number:

2D1

This antibody has been used to optimise flow cytometry-based immunophenotyping of PBMC 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. This antibody was used within a ten-colour staining panel to confirm presence of lymphocytes. The antibody is easy to use and gives clear staining.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human whole blood

Primary Incubation

3µL APC/Fire750-CD45 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

Staining allowed the clear distinction of lymphocytes. Titration optimisations showed the volume of antibody could be reduced from that suggested by the manufacturers. 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

Related Categories

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Summary

The Good

Clear staining of lymphocytes that is useful for flow cytometry analysis of whole blood.

The Bad

The antibody has only been tested for flow cytometry.

The Bottom Line

Easy-to-use pack that can be titrated below manufacturers suggestions and provides clear staining in a multi-colour panel.

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