MHC Class II I-Ab Monoclonal Antibody (AF6-120.1), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Description

• Description: This AF6-120.1.2 monoclonal antibody reacts with the mouse MHC Class II I-Ab alloantigen of H-2b bearing mouse strains, including C57Bl/6 and 129
• This cell surface molecule is involved in antigen presentation to T cells expressing CD3/TCR and CD4
• The AF6-120.1 antibody crossreacts with H-2k and H-2u haplotypes
• However, reactivity to other haplotypes (i.e., d, f, g7, p, q, r, and s) has not been observed
• Applications Reported: This AF6-120.1 antibody has been reported for use in flow cytometric analysis
• Applications Tested: This AF6-120.1 antibody has been tested by flow cytometric analysis of C57Bl/6 splenocytes
• This can be used at less than or equal to 0.5 μg per test
• A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL
• Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test
• It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest
• PerCP-eFluor™ 710 can be used in place of PE-Cy5, PE-Cy5.5 or PerCP-Cy5.5
• PerCP-eFluor™ 710 emits at 710 nm and is excited with the blue laser (488 nm)
• Please make sure that your instrument is capable of detecting this fluorochrome
• For a filter configuration, we recommend using the 685 LP dichroic mirror and 710/40 band pass filter, however the 695/40 band pass filter is an acceptable alternative
• I-Ab (H2-Ab1, histocompatibility 2, class II antigen A, beta 1) is an MHC class II heterodimer molecule of non-covalently associated alpha (31-34 kDa) and beta (26-29 kDa) chains
• Major histocompatibility complex class II antigen presentation requires the participation of lysosomal proteases in two convergent processes
• First, the antigens endocytosed by the antigen presenting cells must be broken down into antigenic peptides
• Second, class II molecules are synthesized with their peptide-binding site blocked by invariant chain (Ii), and they aquire the capacity to bind antigens only after Ii has been degraded in the compartments where peptides reside
• MHC class II molecules present exogenously derived antigen to CD4+ T lymphocytes, which are usually T helper cells
• CD4 interacts with non-polymorphic residues of MHC class II
• H2-Ab1 heterodimer chains alpha (DQA) and beta (DQB) are both anchored in the membrane
• H2-Ab1 plays a central role in the immune system by presenting peptides derived from extracellular proteins
• Class II molecules are expressed in antigen presenting cells (APC: B lymphocytes, dendritic cells, macrophages)
• The beta chain contains 6 exons
• Exon one encodes the leader peptide, exons 2 and 3 encode the two extracellular domains, exon 4 encodes the transmembrane domain and exon 5 encodes the cytoplasmic tail
• Within the DQ molecule both the alpha chain and the beta chain contain the polymorphisms specifying the peptide binding specificities, resulting in up to 4 different molecules
• Typing for polymorphisms is routinely done for bone marrow transplantation
• HLA and MHC antibodies play a significant role in Immunopeptidomics, facilitating the identification and characterization of neoantigens through high-performance liquid chromatography coupled to tandem Mass Spectrometry.