Clone:
REA1031
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
QA1, EA1.2, EA2.1, HLA-6.2, MHC, HLA-E

Extended validation for HLA-E Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA1031
22F6++
736440-
Cells were incubated with an excess of purified unconjugated HLA-E (REA1031) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for HLA-E. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with HLA-E antibodies and with a suitable counterstaining. As a control, HLA-E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for HLA-E. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with HLA-E antibodies and with a suitable counterstaining. As a control, HLA-E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for HLA-E. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with HLA-E antibodies and with a suitable counterstaining. As a control, HLA-E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for HLA-E. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with HLA-E antibodies and with a suitable counterstaining. As a control, HLA-E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for HLA-E. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with HLA-E antibodies and with a suitable counterstaining. As a control, HLA-E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using HLA-E (REA1031). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using HLA-E (REA1031). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using HLA-E (REA1031). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for HLA-E Antibody, anti-human, REAfinity™

Overview

Clone REA1031 recognizes the human HLA-E protein, a member of the human leukocyte antigen family. HLA-E is a non-classical MHC molecule (MHC-Ib) with limited polymorphisms, which is expressed on the cell surface in a heterodimeric complex with β2-microglobulin. HLA-E is ubiquitously expressed and has a specialized role in NK cell recognition. It binds a restricted set of peptides, e.g., derived from other class I leader sequences. The complex is recognized by NKG2A on natural killer (NK) cells. This recognition protects the HLA-E–expressing cell from NK cell killing. Recently HLA-E recognition by a specialised subset of CD8 T cells has been implicated in pathogen recognition as well as autoimmunity.
Additional information: Clone REA1031 displays negligible binding to Fc receptors.

Alternative names

QA1, EA1.2, EA2.1, HLA-6.2, MHC, HLA-E

Detailed product information

Technical specifications

CloneREA1031
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenHLA-E
Alternative names of antigenQA1, EA1.2, EA2.1, HLA-6.2, MHC, HLA-E
Molecular mass of antigen [kDa]38
Entrez Gene ID3133
RRIDAB_2733610, AB_2733750, AB_2733751, AB_2733326, AB_2733327, AB_2732950, AB_2732951, AB_2733490, AB_2733491, AB_2921850, AB_2921831, AB_2922165, AB_2922131, AB_2733609

Resources for HLA-E Antibody, anti-human, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for HLA-E Antibody, anti-human, REAfinity™

Publications

  1. Lu, L. et al. (2008)
    Generation and regulation of CD8
    +
    regulatory T cells.
    Cell. Mol. Immunol. 5(6): 401-406
  2. Jiang, H. et al. (2010)
    HLA-E-restricted regulatory CD8
    +
    T cells are involved in development and control of human autoimmune type 1 diabetes.
    J. Clin. Invest. 120(10): 3641-3650
  3. Pietra, G. et al. (2010)
    The emerging role of HLA-E-restricted CD8
    +
    T lymphocytes in the adaptive immune response to pathogens and tumors.
    J. Biomed. Biotechnol. : 907092

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