Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3664(2)-25] to HIF Prolyl Hydroxylases
  • Suitable for: WB, Flow Cyt, IHC-P, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25]
    See all HIF Prolyl Hydroxylases primary antibodies

  • Description

    Rabbit monoclonal [EPR3664(2)-25] to HIF Prolyl Hydroxylases

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Human cerebrum and lung cancer tissue Flow cyt: HEK-293 cells IP: HEK-293 whole cell lysate WB: HEK-293 and HeLa whole cell lysates

  • 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)

    Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling HIF Prolyl Hydroxylases with ab255733 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human cerebrum (PMID: 17726031, 12163023). The section was incubated with ab255733 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)

    Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling HIF Prolyl Hydroxylases with ab255733 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human lung cancer (PMID: 24382781). The section was incubated with ab255733 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Flow Cytometry - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
    Flow Cytometry - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293 (human embryonic kidney epithelial cell) cells labelling HIF Prolyl Hydroxylases with ab255733 at 1/400 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
    Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)

    HIF Prolyl Hydroxylases was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10μg with ab255733 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab255733 1/1000 dilution (0.4 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

    Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10μg

    Lane 2: ab255733 IP in HEK-293 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255733 in HEK-293 whole cell lysate.

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

    Exposure time: 1 second.

  • Western blot - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
    Western blot - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
    All lanes : Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733) at 1/1000 dilution

    Lane 1 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 57 kDa
    Observed band size: 47 kDa why is the actual band size different from the predicted?


    Exposure time: 20 seconds


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

  • Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
    Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)

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