Tetramer assay - Wikipedia
A tetramer assay (also known as a tetramer stain) is a procedure that uses tetrameric proteins to detect and quantify T cells that are specific for a given antigen within a blood sample. [1] The tetramers used in the assay are made up of four major histocompatibility complex (MHC) molecules, which are found on the surface of most cells in the ...
More tricks with tetramers: a practical guide to staining T cells …
Tetramer staining of T cells is improved by using bright fluorochromes, protein kinase inhibitor treatment and addition of anti-CD8 antibody after tetramer staining.
- tetramers can give a lot of background staining, so you should always gate out B220+ cells and dead cells (i.e. Hoescht negative cells). Also stain with CD4 (I use PETR). - KRN tetramer (GPI/g7) stains in a tight smear. BDC tetramer (2.5mi/g7) gives better separation. - B6 cells should show very minimal tetramer staining (<1%) Materials:
MHC Tetramer Suggested Staining Protocol | BCM
Sometimes, higher intensity tetramer stains are obtained if incubated at room temperature or above, but some surface markers — particularly CD62L— are sensitive to the higher temperature. We recommend you investigate staining at 4°C for …
More tricks with tetramers: a practical guide to staining T
In 2009, we reviewed a number of 'tricks' that could be used to improve this powerful technology. More recent advances have demonstrated the potential benefits of using higher order multimers and of 'boosting' staining by inclusion of an antibody against the pMHC multimer.
More recent advances have demonstrated the potential benefits of using higher order multimers and of ‘boosting’ staining by inclusion of an antibody against the pMHC multimer.
Optimal staining was seen with 2 ug/ml tetramer at 37ºC (figure 1). At 4ºC, very little staining was seen at any tetramer concentration. In a separate experiment, it was found that significant staining took place within 30 mins, but longer incubations led to slight increases (figure 2).
Why Tetramers? | NIH Tetramer Core Facility - Emory University
Antigen-specific T cell responses can be detected by functional assays—e.g. lymphoproliferation assays, cytotoxic T cell assays using chromium release, and cytokine production assays such as the ELISpot assay and intracellular cytokine staining—or by antigen-binding methods.
Tetramer staining of antigen-specific T cells - Purdue University
In our laboratory, tetramer staining is combined with various functional assays, including intracellular cytokine staining and IFN-gamma Elispot after peptide stimulation. We are currently developing technology to produce HLA class II tetramers.
Detection of Antigen-Specific T Cells Using In Situ MHC Tetramer Staining
The development of in situ major histocompatibility complex (MHC) tetramer (IST) staining to detect antigen (Ag)-specific T cells in tissues has radically revolutionized our knowledge of the local cellular immune response to viral and bacterial ...