Anti-HLA E antibody [MEM-E/08] (ab11821)

Reviews (1)Q&A (6)References (7)

Overview

  • Product name
    Anti-HLA E antibody [MEM-E/08]
    See all HLA E primary antibodies
  • Description
    Mouse monoclonal [MEM-E/08] to HLA E
  • Host species
    Mouse
  • Specificity
    The antibody recognized native surface-expressed HLA E, but not denaturated heavy chain of HLA E. The published results revealed that antibody is remarkably specific for HLA E, only with weak cross-reactivity with following classical MHC Class I molecules: HLA A24, B7, B27, B51, B54, C7.
  • Tested applications
    Suitable for: WB, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Bacterially expressed recombinant HLA-E refolded with beta2-microglobulin and peptide.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Purification notes
    Purified from hybridoma culture supernatant by protein A-affinity chromatography.
  • Clonality
    Monoclonal
  • Clone number
    MEM-E/08
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab11821 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
Flow Cyt Use a concentration of 4 µg/ml.

Tested on HLA-E transfected human cell line LCL 721.221

 

 

 

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
IP Use at an assay dependent concentration.

Target

  • Relevance
    HLA 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 localization
    Membrane; 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

    Images

    • Flow Cytometry - Anti-HLA E antibody [MEM-E/08] (ab11821)
      Flow Cytometry - Anti-HLA E antibody [MEM-E/08] (ab11821)This image is courtesy of an anonymous Abreview
      ab11821 staining HLA E in Human K562 cells by Flow Cytometry. The sample was incubated with the primary antibody, (1/50), for 1 hour at 24°C. Ab6785(1/100) was used as secondary antibody.
      Gating Strategy: Isotrope control

      See Abreview

    • Western blot - Anti-HLA E antibody [MEM-E/08] (ab11821)
      Western blot - Anti-HLA E antibody [MEM-E/08] (ab11821)
      Anti-HLA E antibody [MEM-E/08] (ab11821) at 1 µg/ml + Human spleen tissue lysate - total protein (ab29699) at 10 µg

      Secondary
      Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 40 kDa
      Observed band size: 40 kDa
      Additional bands at: 50 kDa, 68 kDa, 73 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 8 minutes

    References

    This product has been referenced in:
    • Wang F  et al. S6K-STING interaction regulates cytosolic DNA-mediated activation of the transcription factor IRF3. Nat Immunol 17:514-522 (2016). WB, IP ; Mouse . Read more (PubMed: 27043414) »
    • Boura JS  et al. Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells. Mol Ther Methods Clin Dev 2014:N/A (2014). Flow Cyt ; Human . Read more (PubMed: 25279386) »
    See all 7 Publications for this product

    Publishing research using ab11821? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-7 of 7 Abreviews or Q&A

    Flow Cytometry abreview for Anti-HLA E antibody [MEM-E/08]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (K562)
    Specification
    K562
    Fixation
    NONE
    Permeabilization
    No
    Gating Strategy
    iostype control
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 20 2010

    Question

    In the data sheet for the HLA-E (MEM-E/08) antibody following crossreactivity is mentioned: The published results revealed that antibody is remarkably specific for HLA E, only with weak cross-reactivity with following classical MHC Class I molecules: HLA A24, B7, B27, B51, B54, C7 Unfortunatly there are no literature references provides to get informed how much cross reactivity is effecting results. Could you send me the literature references?

    Read More
    Answer

    Thank you for your patience in this matter. The originator of ab11821 (HLA E antibody [MEM-E/08]) has provided the following references: Sibilio L. et al., Tissue Antigens 62, 339, 2003. (Abstract 45 from the 3rd Int. Conference on HLA-G, Paris, July 2003). Palmisano GL, Contardi E, Morabito A, Gargaglione V, Ferrara GB, Pist illo MP.: HLA-E surface expression is independent of the availability of HLA class I signal sequence-derived peptides in human tumor cell lines. Hum Immunol. 2005 Jan;66(1):1-12. I hope this helps.

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    Question

    I am trying to titrate the antibody but need the appropriate cells/conditions, because HLA-E is not constitutively expressed on the surface of all cells. I tried fibroblasts and saw some slight internal staining, but only at the 1:12.5 dilution, which is very weak. Once I establish the correct titer to use, I can test the antibody for cross reactivity with rhesus macaque cells, the ultimate goal. How was this antibody tested for application in FACS? What dilution and samples were used?

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    Answer

    This antibody was tested on the human cell line LCL 721.221 (HLA-E transfected), dilution 1:50 - 1:500. This antibody reacts with the native molecule of HLA-E. I hope this helps. Please contact us again if you have any additional questions.

    Read More

    Question

    It is believed that HLA-E is universal expressed on lymphocyte (Human Immunology 1990 (29),131 & Tissue Antigen 1999 (53),292). I used primary aortic endothelial cells and peripheral lymphocytes to test this antibody. Both of them showed negative results. I don't know it is because the expression level is too low to see or other reasons. Did you test this antibody on lymphocyte?

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    Answer

    Thank you for your enquiry. This antibody perfectly works in flow cytometry but as far as we know, this antibody was not tested on lymphocytes. It was tested in FACS with HLA-E transfected human cell line LCL 721.221. Have you tried titrating the antibody? We generally recommend a starting point of 1ug/ml per 1x10^6 cells. However, since we don't have a recommended dilution on the datasheet, it would be important to optimize. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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    Question

    What types of cells has this antibody been tested on in FACS?

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    Answer

    Thank you for your equiry. For testing this antibody on FACS was used human cell line LCL 721.221, HLA-E transfected. Please do not hesitate to contact us if you need anything further.

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    Question

    I wish to use the above antibody for Flow cytometry on HeLa lines, but am unsure at what levels HLA-E is expressed on these cells. Do you have any data on the level of HLA-E expression demonstrated by HeLa cells?

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    Answer

    Thank you for your enquiry and your interest in our products. This antibody perfectly works in flow cytometry but we have not used HeLa cell line for this study. Perhaps this information can be found in some published paper.

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    Question

    I was interested in purchasing MEM-E/08 for flow cytometry. could you confirm whether or not this reagen interacts with HLa-Cw*0401 and send me the reference for the specificty testing of this antibody.

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    Answer

    The antibody appears to be very specific and most likely will not reacts with the allele of question. We will update our on-line datasheet with the cross-reactivity results very soon.

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

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