Staining of peripheral blood from a C57BL/6 mouse (left) or a mouse strain known to express the Eα52-68 peptide:I-Ab complex (right) with Anti-Human/Mouse CD45R (B220) APC (cat. 17-0452) and 0.25 µg of Anti-Mouse Eα52-68 peptide bound to I-Ab Biotin followed by Streptavidin PE (cat. 12-4317). Cells in the lymphocyte gate were used for analysis.
Contents: Anti-Mouse Ea52-68 peptide bound to I-Ab Biotin Catalog Number: 13-5741 Concentration: 0.5 mg/ml Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN3 Storage Conditions: Store at 2-8°C. Do not freeze. Clone: eBioY-Ae (YAe, Y-Ae) Host/Isotype: Mouse IgG2b
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13-5741-81
50 ug
13-5741-82
100 ug
13-5741-85
500 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
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 determinant expressed on a subset of class II I-Ab molecules in strains that also express class II I-Eb. This determinant 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 determinant 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
For research use only, not for diagnostic or therapeutic use. 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 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Murphy DB, Lo D, Rath S, Brinster RL, Flavell RA, Slanetz A, Janeway CA Jr.
A novel MHC class II epitope expressed in thymic medulla but not cortex.
Nature. 1989 Apr 27;338(6218):765-8. (Y-Ae, FC, WB, IHC, PubMed)
Rudensky AYu, Rath S, Preston-Hurlburt P, Murphy DB, Janeway CA Jr.
On the complexity of self.
Nature. 1991 Oct 17;353(6345):660-2. (Y-Ae, FC, WB, PubMed)
Oono T, Fukui Y, Masuko S, Hashimoto O, Ueno T, Sanui T, Inayoshi A, Noda M, Sata M, Sasazuki T.
Organ-specific autoimmunity in mice whose T cell repertoire is shaped by a single antigenic peptide.
J Clin Invest. 2001 Dec;108(11):1589-96. (Y-Ae, FC, IHC, PubMed)
