Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Mouse monoclonal [MEM-E/02] to HLA E
  • Suitable for: Flow Cyt, WB, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-HLA E antibody [MEM-E/02]
    See all HLA E primary antibodies

  • Description

    Mouse monoclonal [MEM-E/02] to HLA E

  • Host species

    Mouse

  • Specificity

    This 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 applications

    Suitable for: Flow Cyt, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein corresponding to Human HLA E.
    Database link: P13747

  • Positive control

    • WB: A549, THP-1 and Jurkat cell lysates. IHC-P: Human tonsil tissue. Flow Cyt: HL60 cells.

  • General notes

    This product has been changed from ascites to tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped 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 buffer

    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Purification notes

    Purified from TCS. Purity >95% by SDS-PAGE.

  • Clonality

    Monoclonal

  • Clone number

    MEM-E/02

  • Myeloma

    unknown

  • Isotype

    IgG1

  • Light chain type

    unknown

  • Research areas

Images

  • Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
    Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
    All lanes : Anti-HLA E antibody [MEM-E/02] (ab2216) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : HLA-E knockout A549 cell lysate
    Lane 3 : THP-1 cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

    Predicted band size: 40 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab2216 observed at 40 kDa. Red - loading control ab181602 observed at 36 kDa.

     ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267081 (knockout cell lysate ab258453) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA E antibody [MEM-E/02] (ab2216)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA E antibody [MEM-E/02] (ab2216)
    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.

  • Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
    Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
    All lanes : Anti-HLA E antibody [MEM-E/02] (ab2216) at 1/500 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : HLA-E knockout A549 cell lysate
    Lane 3 : THP-1 cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

    Predicted band size: 40 kDa
    Observed band size: 40 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab2216 observed at 40 kDa. Red - loading control ab181602 observed at 36 kDa.

     ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267080 (knockout cell lysate ab258452) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
    Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
    All lanes : Anti-HLA E antibody [MEM-E/02] (ab2216) at 1/500 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : HLA-E knockout HEK-293T cell lysate
    Lane 3 : THP-1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 40 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab2216 observed at 40 kDa. Red - loading control ab52901 observed at  kDa.

     ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267231 (knockout cell lysate ab258454) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-HLA E antibody [MEM-E/02] (ab2216)
    Flow Cytometry - Anti-HLA E antibody [MEM-E/02] (ab2216)
    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.

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