HLA-E recombinant antibody

Anti-HLA-E [3D12], Mouse IgG1, kappa

Cat. Num. AAA72554

• Gene Names: HLA-E; MHC; QA1; EA1.2; EA2.1; HLA-6.2
• Reactivity: Human
• Applications: Immunofluorescence, Functional Assay, Flow Cytometry, Functional Assay
• Purity: Protein A affinity purified
• Synonyms: HLA-E, Recombinant Antibody; Anti-HLA-E [3D12], Mouse IgG1, kappa; HLA-E; Anti-HLA-E [3D12]; HLAE; sHLA-E; HLA-6.2; HLA class I histocompatibility antigen alpha chain E; Major histocompatibility complex class I E; MHC; MHC class I antigen E; MHC HLA E alpha 1; MHC HLA E alpha 2.1; HLAB; HLA-B; HLA-C; HLAC; HLA-E recombinant antibody

• Host: Mouse
• Reactivity: Human
• Clonality: Monoclonal
• Isotype: Mouse IgG1 kappa
• Clone Number: 3D12
• Specificity: This antibody binds an extracellular epitope on the human HLA-E, a member of human leukocyte antigen family. This antibody is capable of reacting with free HLA-E heavy chain as well as heavy chain associated with b2-microglobulin and peptide. This antibody is reported to cross react with HLA-B and HLA-C but not with HLA-A, HLA-F or HLA-G. The human HLA-E is a non-classical MHC class 1B molecule involved in immune self-nonself discrimination. HLA-E plays a very important role in cell recognition by NK cells. Peptide-bound HLA-E-B2M heterotrimeric complex primarily functions as a ligand for natural killer (NK) cell inhibitory receptor KLRD1-KLRC1, enabling NK cells to monitor the expression of other MHC class I molecules in healthy cells and to tolerate self.
• Purity/Purification: Protein A affinity purified
• Form/Format: PBS with 0.02% Proclin 300.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for HLA-E recombinant antibody: IF (Immunofluorescence), FA (Functional Assay), FCM/FACS (Flow Cytometry)
• Preparation and Storage: Store at 4 degree C for up to 3 months. For longer storage, aliquot and store at -20 degree C.

FCM/FACS (Flow Cytometry)

(Flowcytometryusinganti-HLA-Eantibody3D12(AAA72554). A431cellswerestainedwiththeanti-unknownspecificityantibodyortherabbitIgGversionof3D12(AAA72554,blueline)atadilutionof1:100for1hatRT.Afterwashing,theboundantibodywasdetectedusingagoatanti-rabbitIgGAlexaFluor488antibodyatadilutionof1:1000,andthecellswereanalyzedusingaFACSCantoflow-cytometer.)

IF (Immunofluorescence)

(ImmunofluorescencestainingofA431cellswithanti-HLA-Eantibody3D12(AAA72554). ImmunofluorescenceanalysisofparaformaldehydefixedA431cellsstainedwiththechimericrabbitIgGversionof3D12(AAA72554)(1:100dilution)for1hfollowedbyAlexaFluor488secondaryantibody(1:1000dilution),showingcytoplasmicstaining.ThenuclearstainisDAPI(blue).Panelsshow,fromleft-right,top-bottom,AAA72554,DAPI,mergedchannelsandanisotypecontrol.TheisotypecontrolwasanunknownspecificityantibodyfollowedbystainingwithAlexaFluor488secondaryantibody.)

WB (Western Blot)

(Western Blotusinganti-HLA-Eantibody3D12(AAA72554). A431cellslysate(35ugproteininRIPAbuffer)wasresolvedviaSDS-PAGE,andthesubsequentblotwasprobedwiththechimericrabbitversionof3D12(AAA72554)at1ug/mlbeforedetectionusingananti-rabbitsecondaryantibody.Aprimaryincubationof1hourwasused,andproteinsweredetectedbychemiluminescence.)

IF (Immunofluorescence)

(ImmunofluorescencestainingofA431cellswithanti-HLA-Eantibody3D12(AAA72554). ImmunofluorescenceanalysisofparaformaldehydefixedA431cellsstainedwiththechimericrabbitIgGversionof3D12(AAA72554)(1:100dilution)for1hfollowedbyAlexaFluor488secondaryantibody(1:1000dilution),showingcytoplasmicstaining.ThenuclearstainisDAPI(blue).Panelsshow,fromleft-right,top-bottom,AAA72554,DAPI,mergedchannelsandanisotypecontrol.TheisotypecontrolwasanunknownspecificityantibodyfollowedbystainingwithAlexaFluor488secondaryantibody.)

Related Product Information for HLA-E recombinant antibody

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).

References

Lee et al. HLA-E surface expression depends on binding of TAP-dependent peptides derived from certain HLA class I signal sequences. J Immunol. 1998 May 15;160(10):4951-60. N/A9590243
Note on publication:
This paper evaluates the regulation of surface expression of HLA-E by signal peptides using antibodies.

NCBI and Uniprot Product Information

• NCBI GI #: 62912479
• NCBI GeneID: 3133
• NCBI Accession #: NP_005507.3
• NCBI GenBank Nucleotide #: NM_005516.5
• UniProt Accession #: P13747; P01889; P10321
• Molecular Weight: 40,157 Da
• NCBI Official Full Name: HLA class I histocompatibility antigen, alpha chain E
• NCBI Official Synonym Full Names: major histocompatibility complex, class I, E
• NCBI Official Symbol: HLA-E
• NCBI Official Synonym Symbols: MHC; QA1; EA1.2; EA2.1; HLA-6.2
• NCBI Protein Information: HLA class I histocompatibility antigen, alpha chain E; HLA class I histocompatibility antigen, E alpha chain; MHC HLA-E alpha-1; MHC HLA-E alpha-2.1; MHC class I antigen E; lymphocyte antigen
• UniProt Protein Name: HLA class I histocompatibility antigen, alpha chain E
• UniProt Gene Name: HLA-E
• UniProt Synonym Gene Names: HLA-6.2; HLAE
• UniProt Entry Name: HLAE_HUMAN