{"id":4566,"date":"2019-04-08T11:29:28","date_gmt":"2019-04-08T11:29:28","guid":{"rendered":"http:\/\/www.caltagmedsystems.co.uk\/information\/?p=4566"},"modified":"2020-03-17T11:05:53","modified_gmt":"2020-03-17T11:05:53","slug":"tips-for-staining-rare-antigens-with-mhc-tetramers","status":"publish","type":"post","link":"https:\/\/www.caltagmedsystems.co.uk\/information\/tips-for-staining-rare-antigens-with-mhc-tetramers\/","title":{"rendered":"Tips for Staining Rare Antigens with MHC Tetramers"},"content":{"rendered":"

\"MBL<\/a>When performing staining with MHC tetramers, there may be certain cases where the staining is unclear. An example could be in certain cases where an antigen-specific T cell population of interest has a low frequency. The following tips discussed in a review in 2015 by Dolton G, Tungatt K, Lloyd A, et al.\u00b9<\/a> may help increase the efficacy of tetramer staining:<\/p>\n

Brighter Fluorochromes<\/h2>\n

Many fluorochromes exist for use with MHC tetramers and monomers. PE, APC, and BV421 can work very well for many common MHC tetramer and monomer applications.<\/p>\n

Use of Protein-Kinase Inhibitor<\/h2>\n

Many T-cell receptors are internalized after exposure to a cognate antigen. This internalization prevents detection of the T-cell receptor using an MHC tetramer. However, by adding a protein-kinase inhibitor such as dasatinib before MHC tetramer staining, the amount of TCR internalization can be reduced and staining intensity increased.<\/p>\n

Use of Anti-Coreceptor Antibody<\/h2>\n

The use of an anti-coreceptor antibody is generally recommended for tetramer staining. However, the clone of the antibody is important to be aware of as certain clones hinder tetramer staining.<\/p>\n

Signal Boosting with Antibodies<\/h2>\n

An increased signal has been found in many instances when using an antibody that targets the tetramer allele. For example, certain HLA-A*02:01 antibodies have been found to increase the staining of HLA-A2 tetramer.<\/p>\n

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Download Free Tetramer Staining Guide<\/a><\/p>\n

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Dolton G, Tungatt K, Lloyd A, et al. (2015) More tricks with tetramers: a practical guide to staining T cells with peptide-MHC multimers. Immunology 146(1):11\u201322. doi: 10.1111\/imm.12499<\/a><\/p>\n

Ready Made MHC Tetramers\u00a0from MBL International<\/h2>\n

MBL International<\/a> has a wide portfolio MHC tetramers for vaccine development, rapid antigen-specific T-cell detection and quantification in the pharmaceutical industry or bioresearch industry. MBL’s Tetramers reduce non-specific binding due to a proprietary mutation in the alpha-3 domain in human MHC class I tetramers.<\/p>\n

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Learn More<\/a><\/p>\n

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Related Blogs<\/h4>\n