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MBL Life Sciences
Anti-mouse CD8 clone choice is critical when mouse MHC I tetramers, as some clones, including 53-6.7, lead to high background staining of all CD8+ T cells, rather than the antigen-specific ones.For best results, clone KT15 in FITC (MBL-K0227-4) is recommended. Additional information can be found in the datasheet that accompanies mouse class I tetramers, as well as the following reference containing a protocol for performing a cross titration series for optimization: Current Protocols in Immunology, MHC-Peptide tetramers to Visualize Antigen-Specific T Cells, Section 17.3.23-24, Supplement 53
T lymphocytesplay a central role in immune system. Total T celland T cell subset counts are measured by detection of various cell surfacemolecules. Enumeration of CD8+antigen-specific T cells requires cognaterecognition of the T cell receptor (TCR) by a class I MHC/peptide complex.This can be done using T-Select MHC class ITetramers which are composed of four MHCclass I molecules each bound to the specificpeptide and conjugated with a fluorescent protein. Thus, T-SelectMHCTetramer assays allow quantitation ofthe total T cell population specificfor a given peptide complexed in a particular MHC molecule. Furthermore,since binding does not depend on functional pathways, thispopulation includes specific CD8+T cells regardless of functionalstatus. Measurements may be performed in whole blood or isolatedlymphocyte/mononuclear cell preparations.In some cases where frequency is low, it may benecessary to perform an in vitrocell expansion. Specific cell stainingis accomplished by incubating the sample with the T-SelectMHC Tetramerreagent, then washing away excess Tetramer. The number of Tetramerpositive lymphocytes is then determined by flow cytometry. This Tetramer reagent comprises murine class I MHC H-2Kband epitope peptide derived from ovalbumin (OVA),and it can detect an H-2Kb-restricted OVA257-264-specific CD8+T cells. The CD8+T cell epitope, OVA257-264, of the OVA model antigen has been a useful tool in immunology. The OT-I transgenic mouse strain carries a TCR transgene specific for the OVA257-264peptide, and it is useful for studying T cell immunology. A Tetramer, which is constructed with the same allele (H-2Kb) of interest and an irrelevant peptide, may be used as a negative control Tetramer. Alternatively, a cell population devoid of Tetramer-positive cells may be used as a negative control.
| Product Specifications | |
| Application | FC |
| Reactivity | Mouse |
| Conjugation | PE |
| Immunogen | OVA |
| Peptide Sequence | SIINFEKL |
| Class | Class I |
| Sequence Location | 257-264 |
| MHC Allele | H-2Kb |
| Documents & Links for H-2Kb OVA Tetramer-SIINFEKL-PE | |
| Datasheet | H-2Kb OVA Tetramer-SIINFEKL-PE Datasheet |
| Documents & Links for H-2Kb OVA Tetramer-SIINFEKL-PE | |
| Datasheet | H-2Kb OVA Tetramer-SIINFEKL-PE Datasheet |
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MBL Life Sciences
Anti-mouse CD8 clone choice is critical when mouse MHC I tetramers, as some clones, including 53-6.7, lead to high background staining of all CD8+ T cells, rather than the antigen-specific ones.For best results, clone KT15 in FITC (MBL-K0227-4) is recommended. Additional information can be found in the datasheet that accompanies mouse class I tetramers, as well as the following reference containing a protocol for performing a cross titration series for optimization: Current Protocols in Immunology, MHC-Peptide tetramers to Visualize Antigen-Specific T Cells, Section 17.3.23-24, Supplement 53
T lymphocytesplay a central role in immune system. Total T celland T cell subset counts are measured by detection of various cell surfacemolecules. Enumeration of CD8+antigen-specific T cells requires cognaterecognition of the T cell receptor (TCR) by a class I MHC/peptide complex.This can be done using T-Select MHC class ITetramers which are composed of four MHCclass I molecules each bound to the specificpeptide and conjugated with a fluorescent protein. Thus, T-SelectMHCTetramer assays allow quantitation ofthe total T cell population specificfor a given peptide complexed in a particular MHC molecule. Furthermore,since binding does not depend on functional pathways, thispopulation includes specific CD8+T cells regardless of functionalstatus. Measurements may be performed in whole blood or isolatedlymphocyte/mononuclear cell preparations.In some cases where frequency is low, it may benecessary to perform an in vitrocell expansion. Specific cell stainingis accomplished by incubating the sample with the T-SelectMHC Tetramerreagent, then washing away excess Tetramer. The number of Tetramerpositive lymphocytes is then determined by flow cytometry. This Tetramer reagent comprises murine class I MHC H-2Kband epitope peptide derived from ovalbumin (OVA),and it can detect an H-2Kb-restricted OVA257-264-specific CD8+T cells. The CD8+T cell epitope, OVA257-264, of the OVA model antigen has been a useful tool in immunology. The OT-I transgenic mouse strain carries a TCR transgene specific for the OVA257-264peptide, and it is useful for studying T cell immunology. A Tetramer, which is constructed with the same allele (H-2Kb) of interest and an irrelevant peptide, may be used as a negative control Tetramer. Alternatively, a cell population devoid of Tetramer-positive cells may be used as a negative control.
| Product Specifications | |
| Application | FC |
| Reactivity | Mouse |
| Conjugation | PE |
| Immunogen | OVA |
| Peptide Sequence | SIINFEKL |
| Class | Class I |
| Sequence Location | 257-264 |
| MHC Allele | H-2Kb |
| Documents & Links for H-2Kb OVA Tetramer-SIINFEKL-PE | |
| Datasheet | H-2Kb OVA Tetramer-SIINFEKL-PE Datasheet |
| Documents & Links for H-2Kb OVA Tetramer-SIINFEKL-PE | |
| Datasheet | H-2Kb OVA Tetramer-SIINFEKL-PE Datasheet |
