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.¹ may help increase the efficacy of tetramer staining:
Brighter Fluorochromes
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.
Use of Protein-Kinase Inhibitor
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.
Use of Anti-Coreceptor Antibody
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.
Signal Boosting with Antibodies
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.
Download Free Tetramer Staining Guide
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):11–22. doi: 10.1111/imm.12499
Ready Made MHC Tetramers from MBL International
MBL International 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.
Related Blogs
- New Alleles added to the MHC Class I Tetramer Range
- Class II MHC Tetramers Available from MBL International
- Non-classical MHC: HLA-E and Qa-1b
- Help! My mouse tetramer stained ALL the CD8+ T cells!
Caltag Medsystems is the UK & Ireland distributor for MBL International. For more information please contact Caltag Medsystems:
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