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    phd2--prolyl-hydroxylase-antibody-epr3660b2-ab133630.pdf

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Cardiovascular Hypoxia Prolyl Hydroxylase
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)

  • Datasheet
  • SDS
Reviews (1) Submit a question References (1)

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Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
  • Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
  • OI-RD Scanning - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
  • Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
  • Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3660(B)(2)] to PHD2 / prolyl hydroxylase
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Human

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Recombinant Human PHD2 / prolyl hydroxylase protein (ab132273)

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Overview

  • Product name

    Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)]
    See all PHD2 / prolyl hydroxylase primary antibodies
  • Description

    Rabbit monoclonal [EPR3660(B)(2)] to PHD2 / prolyl hydroxylase
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    IP
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human PHD2/ prolyl hydroxylase (N terminal). The exact sequence is proprietary.

  • Positive control

    • SH-SY5Y treated with cobalt chloride, SH-SY5Y, and Human adrenal gland lysates
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please 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 long term. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)

    KD = 1.43 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3660(B)(2)
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Hypoxia
    • Prolyl Hydroxylase
    • Epigenetics and Nuclear Signaling
    • Cardiovascular/Immune
    • Hypoxia
    • Prolyl hydroxylase
    • Cancer
    • Invasion/microenvironment
    • Hypoxia
    • Hydroxylases
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia

Associated products

  • Alternative Versions

    • Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (ab232565)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human PHD2 / prolyl hydroxylase protein (ab132273)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133630 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IP
Human
WB
Human
Application Abreviews Notes
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 46 kDa.
IP
1/10 - 1/100.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 46 kDa.
IP
1/10 - 1/100.
  • Application notes
    Is unsuitable for ICC or IHC-P.
  • Target

    • Function

      Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF-1 alpha at 'Pro-402' and 'Pro-564', and HIF-2 alpha. Functions as a cellular oxygen sensor and, under normoxic conditions, targets HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitination complex.
    • Tissue specificity

      According to PubMed:11056053, widely expressed with highest levels in skeletal muscle and heart, moderate levels in pancreas, brain (dopaminergic neurons of adult and fetal substantia nigra) and kidney, and lower levels in lung and liver. According to PubMed:12351678 widely expressed with highest levels in brain, kidney and adrenal gland. Expressed in cardiac myocytes, aortic endothelial cells and coronary artery smooth muscle.
    • Involvement in disease

      Defects in EGLN1 are the cause of erythrocytosis familial type 3 (ECYT3) [MIM:609820]. ECYT3 is an autosomal dominant disorder characterized by increased serum red blood cell mass, elevated serum hemoglobin and hematocrit, and normal serum erythropoietin levels.
    • Sequence similarities

      Contains 1 Fe2OG dioxygenase domain.
      Contains 1 MYND-type zinc finger.
    • Target information above from: UniProt accession Q9GZT9 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 54583 Human
      • Omim: 606425 Human
      • SwissProt: Q9GZT9 Human
      • Unigene: 444450 Human
      • Alternative names

        • C1ORF12 antibody
        • Chromosome 1 Open Reading Frame 12 antibody
        • DKFZp761F179 antibody
        • ECYT 3 antibody
        • ECYT3 antibody
        • Egl 9 family hypoxia inducible factor 1 antibody
        • EGL 9 homolog of C. elegans 1 antibody
        • EGL nine (C.elegans) homolog 1 antibody
        • Egl nine homolog 1 (C. elegans) antibody
        • Egl nine homolog 1 antibody
        • Egl nine like protein 1 antibody
        • EGLN 1 antibody
        • Egln1 antibody
        • EGLN1_HUMAN antibody
        • HIF PH2 antibody
        • HIF Prolyl Hydroxylase 2 antibody
        • HIF-PH2 antibody
        • HIF-prolyl hydroxylase 2 antibody
        • HIFP4H 2 antibody
        • HIFPH2 antibody
        • HPH 2 antibody
        • HPH-2 antibody
        • HPH2 antibody
        • Hypoxia inducible factor prolyl hydroxylase 2 antibody
        • Hypoxia-inducible factor prolyl hydroxylase 2 antibody
        • ORF13 antibody
        • P4H2 antibody
        • PHD 2 antibody
        • PhD2 antibody
        • PNAS 118 antibody
        • PNAS 137 antibody
        • Prolyl Hydroxylase Domain Containing Protein 2 antibody
        • Prolyl hydroxylase domain-containing protein 2 antibody
        • Rat Homolog of SM20 antibody
        • SM 20 antibody
        • SM-20 antibody
        • SM20 antibody
        • Zinc finger MYND domain containing protein 6 antibody
        • ZMYND6 antibody
        see all

      Images

      • Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
        Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)

        Lane 1: Wild-type HAP1 cell lysate (20 µg)
        Lane 2: PHD2 / prolyl hydroxylase knockout HAP1 cell lysate (20 µg)
        Lane 3: HeLa cell lysate (20 µg)
        Lane 4: HepG2 cell lysate (20 µg)
        Lanes 1 - 4: Merged signal (red and green). Green - ab133630 observed at 1X kDa. Red - loading control, ab8245, observed at 37 kDa.

        Unpurified ab133630 was shown to specifically react with PHD2 / prolyl hydroxylase when PHD2 / prolyl hydroxylase knockout samples were used. Wild-type and PHD2 / prolyl hydroxylase knockout samples were subjected to SDS-PAGE. Unpurified ab133630 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW)  preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD)  preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

      • Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
        Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)

        ab133630 (purified) at 1:40 dilution (2µg) immunoprecipitating PHD2 / prolyl hydroxylase in SH-SY5Y treated with 0.1mM cobalt chloride for 8 hours whole cell lysate.
        Lane 1 (input): SH-SY5Y (Human neuroblastoma epithelial cell) treated with 0.1mM cobalt chloride for 8 hours whole cell lysate 10µg
        Lane 2 (+): ab133630 & SH-SY5Y treated with 0.1mM cobalt chloride for 8 hours whole cell lysate
        Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133630 in SH-SY5Y treated with 0.1mM cobalt chloride for 8 hours whole cell lysate
        For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
        Blocking and diluting buffer: 5% NFDM/TBST.

      • OI-RD Scanning - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
        OI-RD Scanning - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
        Equilibrium disassociation constant (KD)
        Learn more about KD

        Click here to learn more about KD
      • Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
        Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
        All lanes : Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630) at 1/5000 dilution (Purified)

        Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates with 5% NFDM/TBST
        Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) treated with 0.1mM cobalt chloride for 8 hours whole cell lysates with 5% NFDM/TBST
        Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

        Predicted band size: 46 kDa
        Observed band size: 46 kDa

      • Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
        Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)

      Protocols

      • Western blot protocols
      • Immunoprecipitation protocols

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (1)

      Publishing research using ab133630? Please let us know so that we can cite the reference in this datasheet.

      ab133630 has been referenced in 1 publication.

      • Bartoszewska S  et al. The hypoxia-inducible miR-429 regulates hypoxia-inducible factor-1a expression in human endothelial cells through a negative feedback loop. FASEB J 29:1467-79 (2015). PubMed: 25550463

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      Western blot abreview for Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)]

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Western blot
      Loading amount
      20 µg
      Gel Running Conditions
      Reduced Denaturing (10%)
      Sample
      Human Cell lysate - whole cell (osteosarcoma cells)
      Specification
      osteosarcoma cells
      Treatment
      lane 1 siRNA for PHD2, lane 2 control, 48h
      Blocking step
      Milk as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 21°C
      Read More

      Dr. Sonia Rocha

      Verified customer

      Submitted Sep 26 2014

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