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Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19656] to HIF-2-alpha
  • Suitable for: IP, WB, ICC/IF
  • Reacts with: Human

Overview

  • Product name

    Anti-HIF-2-alpha antibody [EPR19656]
    See all HIF-2-alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR19656] to HIF-2-alpha

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Whole cell lysate of HeLa cells treated with 1mM deferoxamine (DFO) mesylate salt for 24h ; Whole cell lysate of HeLa cells treated with 100µM CoCl2 for 24h. ICC/IF: HeLa cells treated with DFO mesylate salt (1mM, 24h).

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
    Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
    All lanes : Anti-HIF-2-alpha antibody [EPR19656] (ab207607) at 1/1000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Whole cell lysate of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 1mM deferoxamine (DFO) mesylate salt for 24h

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 96 kDa
    Observed band size: 110 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with the literature, and the expression level can be induced under hypoxia or treated with hypoxia mimetic chemicals such as DFO (PMID: 25587023, 10934146, 23958427 and 21571835).

  • Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
    Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, untreated or treated with deferoxamine mesylate salt (1mM, 24h), labeling HIF-2-alpha with ab207607 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HeLa cells treated with deferoxamine mesylate salt (1mM, 24h).

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
    Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

    HIF-2-alpha was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell treated with 1mM DFO for 24h, cell lysate 10 µg with ab207607 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab207607 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate (10 µg) treated with 1mM DFO for 24h

    Lane 2: ab207607 IP in DFO treated HeLa cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207607 in HeLa cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 75 seconds

     

  • Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
    Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
    All lanes : Anti-HIF-2-alpha antibody [EPR19656] (ab207607) at 1/1000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Whole cell lysate of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 100µM CoCl2 for 24h

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 96 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with the literature, and the expression level can be induced under hypoxia or treated with hypoxia mimetic chemicals such as CoCl2 (PMID: 21571835, 10934146 and 23958427).

  • Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
    Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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