Easily Accessible and Cheap TCR beta Antibody From BioLegend for Flow Cytometry

October 09, 2023

Department of Immunology and Microbiology
Scripps Research
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

PerCP/Cyanine5.5 anti-mouse TCR β chain Antibody

Catalog Number:

109228

Clone Number:

H57-597

The PerCPCy5.5 TCRbeta antibody can be used for identifying T cells within immune cell populations isolated from mouse lungs, spleen, and lymph nodes. The antibody has given good separation at dilution of 1:400 without too many compensation issues. However, I have noticed batch-to-batch effects and more recently, the TCRbeta PerCPCy5.5 antibody I purchased showed very poor staining. I would recommend the TCRbeta APC fire 750 antibodies instead!

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Mouse lung, mediastinal lymphnode, spleens

Primary Incubation

1:400, 30 minutes at 4 degrees C in the dark

Blocking Agent

Fc Shield

Secondary Incubation

BD cytofix/cytoperm; 100 ul for 20 mins at 4 degrees C

Tertiary Incubation

None

Detection

5 laser Aurora Flow cytometer

Results Summary

I used this product to identify TCRbeta+ T cells in mice immunized with antigen. Cells were isolated from lungs, Spleen through digestion or smashing through 70 um filters; RBC lysis was performed using ACK buffer. For lungs, Percoll was used for enrichment of lymphocytes. Cells were then stained with viability dye followed by extra-cellular antibodies such as TCRbeta, CD4, CD8. The T cell pool looks good with this antibody; however, I would recommend the APC-fire 750 conjugated TCRbeta instead for reliability.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Cheap, and good separation.

The Bad

Lot to lot effects and unreliable.

The Bottom Line

A good antibody fluorophore conjugate for TCR beta. However, I would recommend APC-fire 750 conjugated TCR beta for better reliability, depending on your panel set up.

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