Bright and Specific CD8 T Cell Tetramer for Identification of Spike Protein Specific CD8+ T Cells

December 21, 2023

Department of Immunology and Microbiology
Scripps Research
Graduate Student

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Company:

NIH tetramer core facility

Product Name:

S539-546 tetramer for SARS-COV-2 Spike protein

Catalog Number:

Clone Number:

None.

The APC-conjugated S539 Tetramer has been working well for identifying CD8+ T cells that respond to the Spike protein from SARS-CoV-2 in vaccinated B6 mice. I used a PE-conjugated tetramer specific to the Ova peptide as a negative control and saw that the S539 tetramer is highly specific. I use this tetramer at 1:150 dilution and it has given me great separation.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse lung, mediastinal lymphnode, spleens

Primary Incubation

1:150, 1 hour at 4C 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 CD8+ T cells that are specific to S539-546, the immunodominant epitope for T cells in B6 mice. Immune cells were isolated from BALF using 1 ml of 1% BSA in PBS and from lungs through collagenase digestion; RBC lysis was performed using ACK buffer. Cells were then stained with viability dye followed by extracellular antibodies such as CD45, TCR beta, CD4, CD8 along with the S539 tetramer for 1 hour at 4 degrees C. I have not seen any spillover issues with this antibody and fluorophore combination.S539-546: VNFNFNGL

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Bright antibody with negligible spillover on desired cell populations.

The Bad

Takes 6-10 weeks to arrive from the NIH core; order ahead of time.

The Bottom Line

I would highly recommend this tetramer for identification of CD8+ T cells that are responsive to the spike protein peptide from SARS-CoV-2 mice.

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