Overview

  • Product nameAnti-HLA E antibody [MEM-E/02]
    See all HLA E primary antibodies
  • Description
    Mouse monoclonal [MEM-E/02] to HLA E
  • SpecificityThis antibody reacts with the denaturated heavy chain of human HLA-E. It does not cross-react with HLA-A, -B, -C or -G. Specifity of the antibody was confirmed on HLA-G/HLA-E Workshop(Victoria 2002).
  • Tested applicationsSuitable for: Flow Cyt, IHC-Fr, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human, Macaque Monkey
  • Immunogen

    Recombinant denatured heavy chain (Human).

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: 15mM Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity>95% by SDS-PAGE
  • Purification notesPurified from ascities by protein-A affinity chromatography.
  • ClonalityMonoclonal
  • Clone numberMEM-E/02
  • Myelomaunknown
  • IsotypeIgG1
  • Light chain typeunknown
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2216 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 1µg for 106 cells.
IHC-Fr Use at an assay dependent concentration.
WB 1/500 - 1/1000. Predicted molecular weight: 43 kDa. (See Abreviews)
IHC-P Use at an assay dependent concentration.

Target

  • RelevanceHLA E belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. HLA E binds a restricted subset of peptides derived from the leader peptides of other class I molecules.
  • Cellular localizationMembrane; Single-pass type I membrane protein
  • Database links
  • Alternative names
    • HLA class I histocompatibility antigen E alpha chain antibody
    • EA1.2 antibody
    • EA2.1 antibody
    • HLA 6.2 antibody
    • HLA class I histocompatibility antigen alpha chain E antibody
    • HLA class I histocompatibility antigen E alpha chain precursor antibody
    • HLA class I histocompatibility antigen E alpha chain precursor antibody
    • HLA6.2 antibody
    • HLAE antibody
    • Lymphocyte antigen antibody
    • Major histocompatibility complex class I E antibody
    • MHC antibody
    • MHC class I antigen E antibody
    • MHC HLA E alpha 1 antibody
    • MHC HLA E alpha 2.1 antibody
    • QA1 antibody
    see all

Anti-HLA E antibody [MEM-E/02] images

  • IHC image of ab2216 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2216, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab2216 staining HLA E in Human brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was permeabilized with methanol + 1% H2O2. Samples were incubated with primary antibody (1/100 in 4% serum + 1% BSA + 0.01% azide) for 12 hours. A Biotin-conjugated Horse anti-mouse IgG monoclonal (1/100) was used as the secondary antibody.

    See Abreview

  • Anti-HLA E antibody [MEM-E/02] (ab2216) at 1/6000 dilution + whole cell lysate prepared from human endothelial cell line at 100 µg

    Secondary
    HRP conjugated goat anti-mouse polyclonal at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 43 kDa
    Observed band size : 43 kDa


    Exposure time : 20 seconds

    Image courtesy of an anonymous Abreview.

    See Abreview

  • Immunohistochemical analysis of frozen Human adult temporal lobe sections, staining HLA E with ab2216, at 1/20 dilution.

    Arrows denote the location of microglia (M) and neurons (N).
  • Overlay histogram showing HL60 cells stained with ab2216 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2216, 1 µg/1x106 cells for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2 µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-HLA E antibody [MEM-E/02] (ab2216)

This product has been referenced in:
  • Zeestraten EC  et al. Combined analysis of HLA class I, HLA-E and HLA-G predicts prognosis in colon cancer patients. Br J Cancer 110:459-68 (2014). WB ; Human . Read more (PubMed: 24196788) »
  • Engels CC  et al. Immunological subtypes in breast cancer are prognostic for invasive ductal but not for invasive lobular breast carcinoma. Br J Cancer N/A:N/A (2014). Human . Read more (PubMed: 24937677) »

See all 14 Publications for this product

Product Wall

Thank you very much for your interest in our anti-HLA-E antibodies.

To our knowledge, these products have not been tested for blocking. However by participating in our AbTrial program, you can now use our products in an untested applicatio...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (human endothelial cell line)
Loading amount 100 µg
Specification human endothelial cell line
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Apr 25 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (brain)
Specification brain
Fixative none
Permeabilization Yes - Methanol + 1%H2O2
Username

Dr. Pascal Durrenberger

Verified customer

Submitted Nov 17 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (JEG-3, K562)
Loading amount 30 µg
Specification JEG-3, K562
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Apr 23 2010

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (eye)
Specification eye
Fixative Paraformaldehyde
Antigen retrieval step Enzymatic
Blocking step Other as blocking agent for 20 minute(s) · Concentration: 5%
Username

Dr. Luminita Paraoan

Verified customer

Submitted Nov 06 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (uveal melanoma)
Loading amount 40 µg
Specification uveal melanoma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Dr. Luminita Paraoan

Verified customer

Submitted Aug 24 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Uveal Melanoma Biopsy (eye))
Loading amount 30 µg
Specification Uveal Melanoma Biopsy (eye)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Miss. Donna Gray

Verified customer

Submitted May 17 2006

Thank you for your email. Cross-reactivity on our product datasheets refers to what species the antibody has been tested for cross-reactivity with. For example, in the case of ab2216, the antibody has been shown to detect human HLA E. It has not yet be...

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This antibody recognizes the 43 kDa HLA-E heavy chain and does not cross-react with denatured form of HLA-A,-B,-C alleles (no band at 45 kDa observed). The specifity was confirmed at HLA-G/HLA-E. Workshop and publication should appear soon in Human Im...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"