Tetramer Titration
Tetramer Titration - Related Videos
07:24
A Protocol for the Production of KLRG1 Tetramer
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Cited by 2 •
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This protocol describes the production of KLRG1 tetramer, which is a powerful tool for the analysis of KLRG1 ligands.
04:25
A Peptide-MHC-I Tetramer Staining Technique to Analyze SIV-Specific CD8+ Memory T Cells
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This video demonstrates a method for characterizing CD8+ memory T cells specific to simian immunodeficiency virus (SIV) using peptide-major histocompatibility complex class I (pMHC-I) tetramers. A pMHC-I tetramer is a complex of four MHC-I molecules bound to SIV-specific peptides and conjugated to a fluorophore. These tetramers selectively bind to T cell receptors (TCRs) on SIV-specific CD8+ T cells, facilitating the characterization of these cells.
10:17
Introduction to Titration
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2023
Source: Laboratory of Dr. Yee Nee Tan — Agency for Science, Technology, and Research Titration is a common technique used to quantitatively determine the unknown concentration of an identified analyte.1-4 It is also called volumetric analysis, as the measurement of volumes is critical in titration. There are many types of titrations based on the types of reactions they exploit. The most common types are acid-base titrations and redox titrations.5-11 In a typical titration process, a standard...
06:58
n-Butyllithium Titration
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Source: Vy M. Dong and Diane Le, Department of Chemistry, University of California, Irvine, CA This experiment will demonstrate a simple technique to titrate and obtain an accurate concentration of the organolithium reagent, n-butyllithium (n-BuLi). Organolithium reagents are extremely air- and moisture-sensitive and proper care must be taken to maintain the quality of the reagent so that it may be used successfully in a reaction. The n-BuLi titration experiments should be performed regularly...
13:58
Peptide:MHC Tetramer-based Enrichment of Epitope-specific T cells
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Cited by 28 •
2012
This protocol describes the use of peptide:MHC tetramers and magnetic microbeads to isolate low frequency populations of epitope-specific T cells and analyze them by flow cytometry. This method enables the direct study of endogenous T cell populations of interest from in vivo experimental systems.
FAQs
Serial dilution exposes antigen-specific T cells to a range of peptide–MHC tetramer concentrations rather than a single staining condition. Flow cytometry can then reveal how staining intensity and the frequency of tetramer-positive cells change across that range. These patterns help characterize recognition and identify a concentration that provides informative signal for subsequent measurements.
The peptide–MHC component provides the antigenic specificity, while the fluorescent label makes bound tetramers detectable by flow cytometry. Tetramers therefore identify T cells carrying matching receptors and produce measurable staining signals. Examining fluorescence across dilutions helps distinguish changes in antigen-specific detection from background signal and supports evaluation of receptor binding characteristics.
Background signal indicates staining that is not clearly attributable to the intended antigen-specific population. During Tetramer Titration, comparing background with tetramer-positive frequency and staining intensity helps determine whether a condition produces a useful, interpretable signal. This comparison matters because an apparently strong fluorescence result is less informative when background obscures antigen-specific detection.
A typical workflow applies serially diluted fluorescent peptide–MHC tetramers to the T-cell sample, measures staining by flow cytometry, and compares the resulting signal across concentrations. The analysis considers fluorescence intensity, the frequency of tetramer-positive cells, and background signal. Researchers can use these comparisons to select suitable staining conditions for later experiments.
Staining intensity describes the strength of the detected signal, whereas tetramer-positive frequency describes the proportion of cells identified by the reagent. Reviewing both measures across the dilution series provides a broader assessment than either measure alone. Together with background signal, they help compare antigen-specific populations and evaluate receptor binding characteristics in experimental samples.
In infection research, the method supports detection and comparison of pathogen-specific T-cell populations in experimental and clinical samples. It can also help assess receptor binding characteristics and optimize staining conditions before monitoring cellular immune responses. Comparing results across samples or conditions provides a way to study antigen-specific populations using a consistent flow-cytometry-based readout.