Product Information

Contents: Affinity Purified anti-mouse Ea52-68 peptide bound to the I-A(b)/MHC Class II
Catalog Number: 14-5741
Sizes: 50 ug, 100 ug, 500 ug
Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
Clone: eBioY-Ae (YAe, Y-Ae)
Isotype: Mouse IgG2b

Staining of peripheral blood from a mouse strain known to express the I-A(b)/MHC Class II:Ea peptide complex with 0.5 μg of Purified Mouse IgG2b Iso Cntrl (cat. 14-4732) (open histogram) or 0.5 μg of Purified anti-mouse Ea52-68 peptide (eBioY-Ae) (colored histogram) followed by Biotin Anti-Mouse IgG (cat. 13-4013)and SAv-PE (cat. 12-4317). Cells in the lymphocyte gate were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-5741	Affinity Purified anti-mouse Ea52-68 peptide bound to the I-A(b)/MHC Class II	N/A	N/A	FC
13-5741	Biotin anti-mouse Ea52-68 peptide bound to the I-A(b)/MHC Class II	N/A	N/A	FC
11-5741	Fluorescein isothiocyanate (FITC) anti-mouse Ea52-68 peptide bound to the I-A(b)/MHC Class II	488	518	FC
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 Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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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 100 μl blood (or per 1 million cells in 100 μl total staining volume). 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)

Related Products

Cat. 11-4011  FITC Anti-Mouse IgG
Cat. 13-4013  Biotin Anti-Mouse IgG (clone Polyclonal)
Cat. 11-4317  Streptavidin-FITC (Fluorescein isothiocyanate)
Cat. 12-4317  Streptavidin-PE (Phycoerythrin)
Cat. 17-4317  Streptavidin Allophycocyanin (SA-APC)
Cat. 14-4732  Affinity Purified Mouse IgG2b Isotype Control