NIH Tetramer Core Facility
S62-74 epitope CD4+ T cell Tetramer
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The PE conjugated S62 Tetramer has been working really well for identifying CD4+ T cells that respond to the ancestral Spike protein from SARS-CoV-2 in vaccinated B6 mice. I tested the PE-conjugated tetramer at dilutions 1:50, 1:100, 1:200 and 1:400 as well as the unrelated I-A(b) human CLIP 87-101 tetramer. I use this tetramer at 1:400 dilution at room temperature for minimal background and great separation between populations.
Flow cytometry
Mouse lung, mediastinal lymphnode, spleens
1:400, 1 hour at room temp
Fc Shield
Extracellular antibodies with CD8 Tetramer
BD cytofix/cytoperm for 20 min at 4 degrees C
5 laser Aurora Flow cytometer
I used this product to identify CD4+ T cells that are specific to S62-74 (VTWFHAIHVSGTNGT), the immunodominant CD4 T cell 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 CD4 S62 Tetramer for 1 hour at room temperature, followed by extra-cellular antibodies such as CD45, TCRbeta, CD4, CD8 along with the S539 tetramer for 30 minutes at 4C. I have not seen any spillover issues with this antibody and fluorophore combination.
Specific, bright, easy to titrate.
Nothing so far.
I highly recommend this tetramer for identification of Spike specific CD4+ T cells in B6 mice.