Okay CD4-KrO Conjugated Antibody

July 19, 2019

BlueskyReddit
JvGCRC
Nottingham Trent University
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Beckman Coulter

Product Name:

CD4-KrO Conjugated Antibody

Catalog Number:

A96417

Clone Number:

13B8.2

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. CD4 was used in a ten-colour panel to help distinguish T helper cells. It is easy to use.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human whole blood

Primary Incubation

10µL KrO-CD4 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 that works well in combination with CD8, to allow the distinction of major T cell populations. The very dim nature of Krome Orange was acceptable in this specific scenario however I feel it would not work well for a less abundant antigen. Titration optimisations showed we could not reduce the volume of antibody used 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 into a ten-colour panel with no compensation issues.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Easy to use antibody that can distinguish T helper cells.

The Bad

The fluorochrome is very dim so the positive population is close to the negative one.

The Bottom Line

This antibody distinguishes T cell helper cells well but the Krome Orange fluorochrome is less than ideal.

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