FITC Anti-Human CD45RO Antibody

December 10, 2018

JvGCRC
Nottingham Trent University
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

FITC anti-human CD45RO Antibody

Catalog Number:

304204

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. This antibody was used within an eight-colour staining panel for T regulatory cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human whole blood

Primary Incubation

2.5 µL FITC-CD45RO 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 Galios, Kaluza software

Results Summary

Clear staining that allowed the gating of CD45RO+ cells. 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 an eight-colour panel with no compensation issues.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Clear staining and useful for FACS analysis. Titrations showed it was possible to reduce the volume suggested by the manufacturer.

The Bad

The antibody has only been tested for flow cytometry

The Bottom Line

Easy-to-use antibody pack that provided clear and distinct staining of CD45RO+ cells

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