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Mouse MHC class II Tetramer

MHC class II tetramers can be used for direct detection of antigen-specific CD4+ T cells.

MHC slass II

I-Ab MOG35-55 Tetramer-PE


Experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis induced by immunization with MOG35-55 peptide, is a widely used experimental model of autoimmune disease. It has been reported that regulatory T (Treg) cells and Th17 cells are involved in the onset of disease, suggesting that the immune balance of CD4+ T cells is important in EAE pathogenesis.

staining of I-A<sup>b</sup> MOG<sub>35-55</sub> Tetramer-PE

C57BL/6 mice were immunized intraperitoneally twice with 100 nmol of I-Ab-restricted MOG35-55 peptide (MEVGWYRSPFSRVVHLYRNGK, MBL code no. TS-M704-P) and 10 µg cholera toxin (MBL code no. RK-01-511) in complete Freund's adjuvant. Eleven days later, splenocytes were isolated and stained with I-Ab MOG35-55 tetramer-PE (MBL code no. TS-M704-1; Day 0). An aliquot of splenocytes was stimulated with 10µg/mL MOG35-55 peptide for 6 days in vitro and then stained with MHC class II tetramer (Day 6). in vitro stimulation of with I-Ab MOG35-55 resulted in increased tetramer-positive CD4+ T cells from immunized (mouse 1 and 2), but not naïve (mouse 3), mice.

I-Ab OVA323-339 Tetramer-PE


OVA is a model antigen commonly used to study immune responses in mice. OT-II transgenic mice express TCR specific for OVA323-339 epitope (ISQAVHAAHAEINEAGR) in the context of I-Ab and serve as an important tool to study differentiation and activation of CD4 T cells. I-Ab OVA323-339 tetramer can be used to monitor antigen-specific CD4 T cell responses in OT-II mice and in various experimental systems using OT-II cells for adoptive transfer.

Figure 1-1: I-Ab OVA323-339 tetramer staining of freshly isolated OT-II splenocytes
I-Ab OVA323-339 tetramer staining offreshly isolated OT-II splenocytes
Figure 1-2:I-Ab OVA323-339 tetramer staining of peptide-stimulated OT-II splenocytes (Day 6)
I-Ab OVA323-339 tetramer staining of peptide-stimulated OT-II splenocytes (Day 6)

Reactivity of I-Ab OVA323-339 tetramer reagent was assessed using freshly isolated spleen cells from OT-II mice. The majority of CD4 T cells were tetramer positive in mouse 2, but not in mouse 1 (Figure 1-1). In vitro stimulation with specific peptide significantly increased the number of tetramer positive cells (Figure 1-2).

Figure 2: I-Ab OVA323-339 tetramer staining of splenocytes from peptide-immunized mice

I-Ab OVA323-339 tetramer staining of splenocytes from peptide-immunized miceOVA-specific CD4 T cells can also be induced in wild-type mice. C57BL/6 mice were immunized twice IP with a mixture of 100 nmol of OVA323-339 peptide and 10 µg of cholera toxin emulsified with adjuvant.
After 11 days, spleen cells were harvested and cultured with a final concentration of 1 µg/mL OVA323-339 peptide for 8 days. Cells were tested on days 0 and 8 of culture for OVA tetramer reactivity. In mice immunized with OVA323-339 peptide, induction of specific T cells by in vitro peptide stimulation was confirmed using I-Ab OVA323-339 tetramer. I-Ab MOG35-55 tetramer, used as a negative control, showed no specific staining.

Antibody recognizing Mouse I-Ad / OVA323-339 specific TCR

Anti-Mouse TCR DO11.10 (clone KJ1.26) is an antibody that binds to the TCR specific for OVA323-339 presented in the context of I-Ad. This antibody can be used to identify OVA323-339-specific CD4 T cells derived from I-Ad mice. OVA323-339 peptide (ISQAVHAAHAEINEAGR, MBL code no. TS-M703-P) is often used as a helper peptide for induction of antigen-specific CTL in I-Ad-expressing mice.

Flow cytometry with Anti-mouse TCR DO11.10 mAb

Antibody recognizing Mouse I-Ad / OVA323-339 specific TCR

I-A alleles of mouse strains

I-A allele
Mouse strains

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