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What is a Multimer in IHC?

What is a Multimer in IHC?

MHC multimers are oligomeric forms of MHC molecules, designed to identify and isolate T-cells with high affinity to specific antigens amid a large group of unrelated T-cells.

What is MHC staining?

In situ MHC-tetramer staining (IST) is a technique to visualize T cells that are specific for antigens of interest in tissues. In combination with immunohistochemistry (IHC), IST can determine the abundance, location, and phenotype of antigen-specific CD8 and CD4 T cells in tissues.

What is a Dextramer?

Dextramer® reagents were designed to help researchers and clinicians worldwide to obtain high-quality results. They contain multiple binding sites and numerous fluorophores to boost avidity in binding antigen-specific T cells to isolate populations even when these are rare or have low-affinity receptors.

What are multimeric proteins?

Descriptive of a protein composed of several subunits.

How do I make MHC tetramers?

MHC tetramers are produced by mixing enzymatically biotinylated MHC fusions to a BirA substrate peptide (BSP) (Schatz, 1993) with fluorescent conjugates of streptavidin or avidin (Altman et al., 1996); the MHC molecules used for tetramer preparations are homogeneous with respect to the peptide ligand that occupies the …

How do MHC tetramers work?

MHC tetramer technology is based on the ability of MHC-peptide complexes to recognize the antigen-specific T cells at a single cell level. This breakthrough technology enables researchers to precisely measure targeted T-cell responses in infectious diseases, cancer, and autoimmune diseases.

What is peptide MHC?

Abstract. Major histocompatibility complex (MHC) antigens bind peptides of diverse sequences with high affinity. They do this in order to generate maximal immunological protection by covering the spectrum of peptides that may be seen by a host over the course of its lifetime.

What does Multimeric mean?

multimeric (comparative more multimeric, superlative most multimeric) (biochemistry) Describing a protein that has multiple polypeptide chains.

What is the subunit of a multimeric protein?

Single chains of amino acids that are the units of multimeric PROTEINS. Multimeric proteins can be composed of identical or non-identical subunits. One or more monomeric subunits may compose a protomer which itself is a subunit structure of a larger assembly.

What are MHC tetramers?

MHC tetramers are complexes of 4 MHC molecules (Major Histocompatibility Complex) wich are associated with a specific peptide and bound to a fluorochrome. Class I tetramers bind to a distinct set of T cell receptors (TRCs) on a subset of T CD8+ cells, Class II tetramers bind to a distinct population of T CD4+ cells.

What are MHC class I tetramers?

Preparation of Class I MHC Tetramers

MHC Tetramers are complexes of four MHC molecules, associated with a specific peptide and bound to a fluorochrome. The T cell surface CD8 enhances T cell antigen recognition by binding to HLA class I molecules.

What is the use of tetramers?

Tetramers/multimers have been used by researchers to identify and quantify antigen-specific T cells from patient samples, which is especially useful when determining the effectiveness of a vaccine3.

What is the difference between MHC I and MHC II?

The main difference between MHC class 1 and 2 is that MHC class 1 molecules present antigens to cytotoxic T cells with CD8+ receptors whereas MHC class 2 molecules present antigens to helper T cells with CD4+ receptors.

Why does MHC bind to peptides?

What is Multimeric protein?

When multiple copies of a polypeptide encoded by a gene form a complex, this protein structure is referred to as a multimer.

What is a multimeric enzyme?

Many enzymes consisting of tetramers or higher order multimers are made from two or more pairs of such coupled sites. These enzymes can then also display positive cooperativity towards the same ligand.

What is the smallest subunit of proteins?

Proteins are made up of hundreds or thousands of smaller units called amino acids, which are attached to one another in long chains. There are 20 different types of amino acids that can be combined to make a protein.

How are MHC tetramers made?

The enzyme BirA is then added to biotinylate the complex, adding a biotin molecule to each MHC monomer that is formed. In the presence of streptavidin which has four biotin binding sites per molecule, four MHC monomers are joined together to form a tetramer.

What are T cell tetramers?

The tetramers are labeled with a fluorophore, allowing tetramer-bound T-cells to be analyzed with flow cytometry.

Tetramer assay
Purpose uses tetrameric proteins to detect and quantify T cells

What is the role of MHC 1 and MHC 2?

Major histocompatibility complex (MHC) class I and class II proteins play a pivotal role in the adaptive branch of the immune system. Both classes of proteins share the task of presenting peptides on the cell surface for recognition by T cells.

Where are MHC 1 and 2 found?

There are two major types of MHC protein molecules—class I and class II. Class I MHC molecules span the membrane of almost every cell in an organism, while class II molecules are restricted to cells of the immune system called macrophages and lymphocytes.

What is MHC restriction of T cells?

MHC restriction refers to the fact that mature αβT cells recognize either lipids or peptides of antigenic proteins that are bound to MHC as peptide-MHC (pMHC) complexes, but essentially nothing else.

What are the 2 subunits of ribosomes?

Ribosomes contain two different subunits, both of which are required for translation. The small subunit (“40S” in eukaryotes) decodes the genetic message and the large subunit (“60S” in eukaryotes) catalyzes peptide bond formation.

What are the three bases of mRNA called?

The mRNA bases are grouped into sets of three, called codons. Each codon has a complementary set of bases, called an anticodon. Anticodons are a part of transfer RNA (tRNA) molecules.

What is the difference between MHC class 1 and MHC Class 2?

MHC class 1 molecules present endogenous antigens to cytotoxic T cells. MHC class 2 molecules present exogenous antigens to helper T cells. Thus, the main difference between MHC class 1 and 2 molecules is the type of antigens presented by each type of MHC molecules.