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使用敲除细胞株进行验证

Anti-HLA E抗体[MEM-E/02] (ab2216)

Reviews (6)Q&A (3)References (16)
Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
  • Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
  • Western blot - Anti-HLA E antibody [MEM-E/02] (ab2216)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA E antibody [MEM-E/02] (ab2216)
  • Flow Cytometry - 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

概述

  • 产品名称

    Anti-HLA E抗体[MEM-E/02]
    参阅全部 HLA E 一抗

  • 描述

    小鼠单克隆抗体[MEM-E/02] to HLA E

  • 宿主

    Mouse

  • 特异性

    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).

  • 经测试应用

    适用于: Flow Cyt, WB, IHC-Pmore details

  • 种属反应性

    与反应: Human

  • 免疫原

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

  • 阳性对照

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

  • 常规说明

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid

  • 存放说明

    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.

  • 存储溶液

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

  • 纯度

    Protein A purified

  • 纯化说明

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

  • 克隆

    单克隆

  • 克隆编号

    MEM-E/02

  • 骨髓瘤

    unknown

  • 同种型

    IgG1

  • 轻链类型

    unknown

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab2216于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB (4)
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.
IHC-P (1)
Use a concentration of 5 - 10 µg/ml.
说明
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.
IHC-P
Use a concentration of 5 - 10 µg/ml.

靶标

  • 相关性

    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.

  • 细胞定位

    Membrane; Single-pass type I membrane protein

  • 数据库链接

    • 别名

      • 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

    图片

    • 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.

    • 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.

    • 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.

    文献 (16)

    发表研究结果有使用 ab2216？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab2216 被引用在 16 文献中.

    • 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 . 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 . PubMed: 24937677
    • Reimers MS  et al. Prognostic value of HLA class I, HLA-E, HLA-G and Tregs in rectal cancer: a retrospective cohort study. BMC Cancer 14:486 (2014). IHC-P ; Human . PubMed: 24997850
    • Shwetank  et al. Inhibition of ERK and proliferation in NK cell lines by soluble HLA-E released from Japanese encephalitis virus infected cells. Immunol Lett 162:94-100 (2014). PubMed: 25086398
    • Zaguia F  et al. Cytotoxic NKG2C+ CD4 T cells target oligodendrocytes in multiple sclerosis. J Immunol 190:2510-8 (2013). Flow Cyt ; Human . PubMed: 23396942
    View all Publications for this product

    客户评价及客户问答

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    1-9 of 9 Abreviews or Q&A

    Western blot abreview for Anti-HLA E antibody [MEM-E/02]

     Excellent
    Abreviews
    abreview image
    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%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Apr 25 2011

    Immunohistochemistry (Frozen sections) abreview for Anti-HLA E antibody [MEM-E/02]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (brain)
    Specification
    brain
    Fixative
    none
    Permeabilization
    Yes - Methanol + 1%H2O2
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Pascal Durrenberger

    Verified customer

    提交于 Nov 17 2010

    Western blot abreview for Anti-HLA E antibody [MEM-E/02]

     Average
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Apr 23 2010

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HLA E antibody [MEM-E/02]

     Below Average
    Abreviews
    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%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Luminita Paraoan

    Verified customer

    提交于 Nov 06 2006

    Western blot abreview for Anti-HLA E antibody [MEM-E/02]

     Good
    Abreviews
    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%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Luminita Paraoan

    Verified customer

    提交于 Aug 24 2006

    Western blot abreview for Anti-HLA E antibody [MEM-E/02]

     Good
    Abreviews
    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%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Miss. Donna Gray

    Verified customer

    提交于 May 17 2006

    Question

    Dear Sirs and Madams, unfortunately I couldn't receive the information I was looking for by calling your hotline, so I would like to ask you this way if you have any information about the blocking abilities of your anti HLA-E antibodies. I also didn't see it anywhere online so I am hoping you could help me choose the best antibody for this task. Thank you very much in advance,

    Read More
    Answer

    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 application or species without financial risk.

    Simply follow these easy steps below to take part in our AbTrial Program:

    1. Reply to this e-mail to let me know that you would like to proceed and test one or more of these products.

    2. I will email you an inactive personal discount code for the value of the product.

    3. Purchase and test the product at regular price.

    4. https://www.abcam.com/abreviews with your results. Note your AbTrial discount code in the additional notes section.

    5. Once the Abreview is submitted, the discount code will become active.

    6. Apply your discount code on your next order to receive that value off.

    More information including the terms and conditions about this program may be found at: https://www.abcam.com/AbTrial

    Read More

    Question

    I am looking for anti-HLA-E antibodies for FACS and Western. I found 4 anti-HLA-E antibodies from your website and I have some technical questions about them, such as, what does "cross-reacts with human" mean on your data sheet when it states "it does not cross-react with HLA-A,-B, -C or -G", and for the other three anti-HLA-E antibodies how serious the cross-reactive problem is. Do you have a technical service number for customers in the US? For ordering, do you have special discount for a bulk purchasing?

    Read More
    Answer

    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 been determined as to whether the antibody will detect mouse HLA E, for instance. All the information, including cross-reactivity, that we have regarding our antibodies is located on the online datasheets. We do have discounts for bulk orders - ordering more than 2 vials of the same antibody. If you have any additional questions, please do let me know.

    Read More

    Question

    I have used the HLA-E antibody for Western blotting and gotten two seemingly very specific bands on my film. What are the molecular weight of the HLA-E part(s) that the antibody regognises? Many thanks. Best Regards

    Read More
    Answer

    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 Immunology.

    Read More

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    For licensing inquiries, please contact partnerships@abcam.com