Description: The Y-Ae antibody reacts with a class II major histocompatibility complex (MHC) self Ea peptide (peptide 52-68) bound to I-Ab molecules. The Y-Ae antibody detects a determit expressed on a subset of class II I-Ab molecules in strains that also express class II I-Eb. This determit is expressed on peripheral B cells and on cells in the thymic medulla, but not thymic cortical epithelium. In addition to B cells, the Y-Ae determit is expressed at comparable levels on other antigen presenting cells, including macrophages and dendritic cells. The antibody does not react with invariant chain-associated class II MHC complexes. The reactivity of the Y-Ae antibody is MHC-restricted. The Y-Ae antibody was generated by immunization of [B10.MBR x B10.D2]F1 recipients with B10A(5R) LPS blasts. In this strain combination, there is incompatibility for I-Ab and I-Eb molecules. The antibody reacts with 10-15% of surface I-Ab molecules in strains that also express surface I-Eb molecules (e.g., strains B10.A(5R) and B10.A(3R)). The antibody does not react with I-A molecules in strains that have a non-functional I-E (a chain) gene (e.g., strains B6 and B10), nor with mutant I-A molecules (e.g., (B6.c-H-2bm-12 x A.TFR5)F1), nor with any other strain tested. The Y-Ae antibody recognizes an Ea peptide (Ea 52-68) bound to I-Ab molecules, but not to Ea 52-68 peptide bound to I-Ak or I-Abm-12 molecules.Applications Reported: This eBioY-Ae (YAe, Y-Ae) antibody has been reported for use in flow cytometric analysis. Applications Tested: This eBioY-Ae (YAe, Y-Ae) antibody has been tested by flow cytometric analysis of a mouse strain known to express the I-A(b)/MHC Class II:Ea peptide complex. 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. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 μm post-manufacturing filtered. MHC class II molecules are heterodimers 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.
|Ea52-68 peptide bound to I-Ab|
|PBS with 0.09% sodium azide; pH 7.2|
|eBioY-Ae (YAe, Y-Ae)|
For Research Use Only.
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