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Rabbit Recombinant Monoclonal PHD2 / prolyl hydroxylase antibody. Suitable for IP, WB and reacts with Human samples. Cited in 3 publications.


Images

Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (AB133630), expandable thumbnail
  • Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (AB133630), expandable thumbnail
  • OI-RD Scanning - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (AB133630), expandable thumbnail
  • Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (AB133630), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPWBIHC-P
Human
Not recommended
Tested
Tested
Not recommended

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/10 - 1/100
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000 - 1/10000
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

3 products for Alternative Product

Target data

Function

Cellular oxygen sensor that catalyzes, under normoxic conditions, the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A. Also hydroxylates HIF2A. Has a preference for the CODD site for both HIF1A and HIF1B. Hydroxylated HIFs are then targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. Under hypoxic conditions, the hydroxylation reaction is attenuated allowing HIFs to escape degradation resulting in their translocation to the nucleus, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. EGLN1 is the most important isozyme under normoxia and, through regulating the stability of HIF1, involved in various hypoxia-influenced processes such as angiogenesis in retinal and cardiac functionality. Target proteins are preferentially recognized via a LXXLAP motif.

Alternative names

C1orf12, PNAS-118, PNAS-137, EGLN1, Egl nine homolog 1, Hypoxia-inducible factor prolyl hydroxylase 2, Prolyl hydroxylase domain-containing protein 2, SM-20, HIF-PH2, HIF-prolyl hydroxylase 2, HPH-2, PHD2

Recommended products

Rabbit Recombinant Monoclonal PHD2 / prolyl hydroxylase antibody. Suitable for IP, WB and reacts with Human samples. Cited in 3 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR3660(B)(2)
Purification technique
Affinity purification Protein A
Dissociation constant
1.43 x 10-11 M
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

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.

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

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The protein PHD2 also known as prolyl hydroxylase 3 EGLN1 or 366G is a member of the prolyl hydroxylase family with a molecular mass of approximately 46 kDa. PHD2 is expressed in a variety of tissues particularly those in the human body's oxygen-sensing cellular machinery. This protein functions mechanically by adding hydroxyl groups to proline residues on its target proteins an enzymatic activity essential for its role in cellular oxygen sensing.

Biological function summary

This hydroxylation process influences the stability of hypoxia-inducible factor (HIF) by marking it for degradation under normal oxygen levels. PHD2 is a monomeric enzyme but its hydroxylase activity does not require association with other proteins to function. By regulating HIF PHD2 plays a critical role in cellular responses to oxygen availability controlling genes involved in processes such as angiogenesis metabolism and cell survival.

Pathways

PHD2 participates in the HIF signaling pathway and oxygen homeostasis pathways. The regulation of HIF by PHD2 occurs alongside PHD1 and PHD3 which have similar hydroxylase activities. Importantly PHD2 protects cells from hypoxia-related stress by ensuring the degradation of HIF-α subunits under normoxic conditions thereby maintaining cellular homeostasis and proper metabolic function.

Associated diseases and disorders

PHD2 has connections with conditions such as cancer and chronic kidney disease. Its role in cancer is linked to its regulation of HIF where dysregulation can lead to abnormal cell growth and angiogenesis. In chronic kidney disease the insights into PHD2's function offer potential therapeutic targets particularly since HIF stabilization can ameliorate anemia frequently associated with this condition. Interactions between PHD2 and other proteins in these disease pathways make it a significant focal point for research aimed at developing new treatments.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

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

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

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

    Predicted band size: 46 kDa

    Observed band size: 46 kDa

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

    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, Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

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

    Predicted band size: 46 kDa

  • OI-RD Scanning - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630), expandable thumbnail

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

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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

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

    PHD2 / prolyl hydroxylase Western blot staining using rabbit Anti-PHD2 / prolyl hydroxylase antibody

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

    All lanes: Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630) at 1/5000 dilution

    Lane 1: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates at 20 µg

    Lane 2: SH-SY5Y (Human neuroblastoma epithelial cell) treated with 0.1mM cobalt chloride for 8 hours whole cell lysates at 20 µg

    Lane 3: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

    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

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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