Clear Staining with This CD28 Antibody from BioLegend

October 08, 2019

JvGCRC
Nottingham Trent University
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

APC anti-human CD28 Antibody

Catalog Number:

302911

Clone Number:

CD28.2

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. CD28 was used in a ten-colour panel to assess expression in 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

2µL APC-CD28 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 CD28 expression on T cell populations. Titration optimisations showed the volume of antibody used could be reduced 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

Related Categories

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Summary

The Good

Clear staining of CD28+ 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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