AB232565

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

RabMAb
Recombinant
KO Validated
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(1 Publication)

Rabbit Recombinant Monoclonal PHD2 / prolyl hydroxylase antibody. Carrier free. Suitable for IP, WB and reacts with Human samples. Cited in 1 publication.

View 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

3 Images

Lab

Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (AB232565)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133630).

All lanes:

Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (<a href='/en-us/products/primary-antibodies/phd2-prolyl-hydroxylase-antibody-epr3660b2-ab133630'>ab133630</a>)

Predicted band size: 46 kDa

Observed band size: 46 kDa

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Supplier Data

Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (AB232565)

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.
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. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133630).

All lanes:

Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (ab232565)

Predicted band size: 46 kDa

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OI-RD Scanning - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (AB232565)

OI-RD Scanning

Unknown

OI-RD Scanning - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (AB232565)

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.

Unconjugated

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

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR3660(B)(2)

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, IP

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" } } }

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Product details

ab232565 is the carrier-free version of ab133630.

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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Supplementary information

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

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.

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.

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

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.

See full target information EGLN1

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

iScience 25:104262 PubMed35521516

2022

Neonatal lung-derived SSEA-1 cells exhibited distinct stem/progenitor characteristics and organoid developmental potential.

Applications

Unspecified application

Species

Unspecified reactive species

Chien-Chia Liao,Chiao-Juno Chiu,Yao-Hsu Yang,Bor-Luen Chiang

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

For licensing inquiries, please contact partnerships@abcam.com

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

Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (AB232565)

Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (AB232565)

OI-RD Scanning - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] - BSA and Azide free (AB232565)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Alternative Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Complementary Products

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

Product promise