Predictions tools like PSSMHCpan, NetMHC-4.0, NetMHC-3.0 and PickPocket are commonly used to predict candidate binding affinities.
Wang, Wenfeng, et al 2023, selected the top 15 top peptides for self-antigen ( ce (NP_001157643.1) for their study. By using the QuickSwitch™ kit (TB-7400-K1), they successfully switched six peptides to the tetramer complex with an efficiency of over 75%. These tetramers stained for CD8+ T cells that specifically bound to the Myh6-peptide-tetramer complex, in heart and mediastinal lymph node samples. A negative peptide (H-2Kb Negative Tetramer -SIYRYYGL-PE, (TS-M008-1) was used for controlling non-specific binding. Overall, the results revealed significant changes in the gene expression of lymphatic endothelial cells (LECs) after myocardial infarction (MI), indicating potential effects on lymphangiogenesis and immunomodulation (1).
MBL International offers QuickSwitch™ Custom Tetramers kit for you to create your own custom tetramer. This kit is supplied with a tetramer bound with an irrelevant peptide. After adding a peptide exchange factor and your peptide of interest, peptide exchange will occur if there is binding affinity of the peptide with the MHC molecule. By using the QUANT kit, you can quantify the amount of exchange that occurred, thereby gathering binding affinity information for the MHC/peptide complex. This kit can be used to create custom tetramers, or for screening peptides, all in one day. QuickSwitch™ is an assay system that allows discrimination of MHC binding from non-binding peptides.
In addition, MBLI negative controls provide for accurate monitoring of immune response. For example, H-2Kb tetramers specific for either β-gal or TRP2 have been used as negative controls in experiments using splenocytes from OT-I mice transgenic for the T cell receptor that binds specifically to H-2Kb OVA (SIINFEKL). For proper gate placement, the full panel of antibodies used for the experimental tetramer stain must be included in the control tetramer stain to account for fluorescent spillover contributed by fluorochromes from other channels of the flow cytometer. In experiments involving immunised or treated mice, often cells from a naïve mouse, stained with the same tetramer/antibody panel as the experimental mouse cells, is an appropriate negative control.
| Product Code | Product Name |
| TB-7400-K1 | QuickSwitch™ Quant H-2 Kb Tetramer Kit-PE |
| TS-M008-1 | T-Select H-2Kb Negative Tetramer-SIYRYYGL-PE |
REFERENCES:
- Wang, Wenfeng, et al. “Lymphatic endothelial transcription factor Tbx1 promotes an immunosuppressive microenvironment to facilitate post-myocardial infarction repair.” Immunity (2023).
Information provided by MBL.
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