Anti-HLA-E [3D12]

• Catalog Number: orb2645782
• Category: Antibodies
• Description: Anti-HLA-E [3D12]
• Target: HLA-E
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human
• Concentration: 1 mg/ml
• Buffer/Preservatives: PBS with 0.02% Proclin 300.
• Immunogen: The original antibody was generated by immunizing HLA-B27 transgenic mice with recombinant human HLA-E purified from AEH cells (Lymphoblastoid cell line LCL 721.221 cells transfected with HLA-E gene).
• UniProt ID: P10321, P01889, P13747
• Tested applications: FA, FC, IF
• Clone Number: 3D12
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: HLAE; sHLA-E; HLA-6.2; HLA class I histocompatibil
• Note: For research use only

Flow cytometry using anti-HLA-E antibody 3D12. A431 cells were stained with the anti-unknown specificity antibody (orb256458; isotype control, black line) or the rabbit IgG version of 3D12 (orb2645782, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000, and the cells were analyzed using a FACSCanto flow-cytometer.

Immunofluorescence staining of A431 cells with anti-HLA-E antibody 3D12. Immunofluorescence analysis of paraformaldehyde fixed A431 cells stained with the chimeric rabbit IgG version of 3D12 (orb2645782) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, orb2645782, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (orb256458) followed by staining with Alexa Fluor® 488 secondary antibody.

Western blot using anti-HLA-E antibody 3D12. A431 cells lysate (35 µg protein in RIPA buffer) was resolved via SDS-PAGE, and the subsequent blot was probed with the chimeric rabbit version of 3D12 (orb2645782) at 1 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and proteins were detected by chemiluminescence.