Anti-PHD3 antibody [EPR17869] (ab184714)

Rabbit Recombinant Monoclonal PHD3 antibody. Suitable for IP, WB, ICC/IF and reacts with Mouse, Rat, Human samples. Cited in 4 publications.

Be the first to review this product! Submit a review

Images

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2
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

	IP	WB	ICC/IF
Human	Expected	Tested	Tested
Mouse	Tested	Tested	Expected
Rat	Expected	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/70	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes -
Species Rat	Dilution info 1/2000	Notes -
Species Human	Dilution info 1/2000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/250	Notes -
Species Human	Dilution info 1/250	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

See full target information EGLN3

Function

Prolyl hydroxylase that mediates hydroxylation of proline residues in target proteins, such as PKM, TELO2, ATF4 and HIF1A (PubMed:19584355, PubMed:20978507, PubMed:21483450, PubMed:21575608, PubMed:21620138, PubMed:22797300). Target proteins are preferentially recognized via a LXXLAP motif. Cellular oxygen sensor that catalyzes, under normoxic conditions, the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins (PubMed:11595184, PubMed:12181324). Hydroxylates a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A (PubMed:11595184, PubMed:12181324). Also hydroxylates HIF2A (PubMed:11595184, PubMed:12181324). Has a preference for the CODD site for both HIF1A and HIF2A (PubMed:11595184, PubMed:12181324). Hydroxylation on the NODD site by EGLN3 appears to require prior hydroxylation on the CODD site (PubMed:11595184, PubMed:12181324). Hydroxylated HIFs are then targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex (PubMed:11595184, PubMed:12181324). 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 (PubMed:11595184, PubMed:12181324). ELGN3 is the most important isozyme in limiting physiological activation of HIFs (particularly HIF2A) in hypoxia. Also hydroxylates PKM in hypoxia, limiting glycolysis (PubMed:21483450, PubMed:21620138). Under normoxia, hydroxylates and regulates the stability of ADRB2 (PubMed:19584355). Regulator of cardiomyocyte and neuronal apoptosis. In cardiomyocytes, inhibits the anti-apoptotic effect of BCL2 by disrupting the BAX-BCL2 complex (PubMed:20849813). In neurons, has a NGF-induced proapoptotic effect, probably through regulating CASP3 activity (PubMed:16098468). Also essential for hypoxic regulation of neutrophilic inflammation (PubMed:21317538). Plays a crucial role in DNA damage response (DDR) by hydroxylating TELO2, promoting its interaction with ATR which is required for activation of the ATR/CHK1/p53 pathway (PubMed:22797300). Also mediates hydroxylation of ATF4, leading to decreased protein stability of ATF4 (Probable).

Alternative names

Prolyl hydroxylase EGLN3, Egl nine homolog 3, HPH-1, Hypoxia-inducible factor prolyl hydroxylase 3, Prolyl hydroxylase domain-containing protein 3, HIF-PH3, HIF-prolyl hydroxylase 3, HPH-3, PHD3, EGLN3

Recommended products

Rabbit Recombinant Monoclonal PHD3 antibody. Suitable for IP, WB, ICC/IF and reacts with Mouse, Rat, Human samples. Cited in 4 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2
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.
Clone number: EPR17869
Purification technique: Affinity purification Protein A
Concentration

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

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 PHD3 protein also known as EGLN3 or Prolyl Hydroxylase Domain-Containing Protein 3 functions mechanically to regulate oxygen homeostasis in cells. It catalyzes the hydroxylation of proline residues on hypoxia-inducible transcription factors (HIFs). The molecular mass of PHD3 is approximately 27 kDa. PHD3 expresses in a variety of tissues notably in the heart brain and skeletal muscles. Its expression often occurs in response to hypoxic conditions reflecting its role in oxygen sensing and adaptation to change in oxygen levels.

Biological function summary

The PHD3 protein plays an essential role in regulating the degradation of HIFs preventing their accumulation under normoxic conditions. It is part of a larger complex which includes oxygen iron and 2-oxoglutarate facilitating its hydroxylase activity. Hydroxylation of HIFs by PHD3 marks them for degradation via the ubiquitin-proteasome pathway preventing HIFs from activating genes related to erythropoiesis angiogenesis and cellular metabolism adaptation to hypoxia. Through these actions PHD3 helps maintain cellular oxygen homeostasis and metabolic balance.

Pathways

PHD3 is integral to the HIF signaling pathway and the cellular response to hypoxia. Its interaction with HIF-1α and HIF-2α is important in this context dictating the stability and activity of these transcription factors under varying oxygen levels. PHD3 also associates with other prolyl hydroxylases such as PHD1 and PHD2 coordinating the regulation of HIFs collectively across different cell types and conditions. These interactions contribute to the modulation of gene expression in response to hypoxic stress.

Associated diseases and disorders

Aberrant PHD3 activity links to cancer and ischemic diseases. In cancer altered PHD3 expression affects tumor growth and metastasis by disrupting normal oxygen sensing allowing cancer cells to adapt to low-oxygen environments. Moreover PHD3's interaction with proteins like HIF-1α and HIF-2α plays a role in the pathological angiogenesis seen in certain cancer types. In ischemic diseases improper regulation by PHD3 might impede normal tissue responses to reduced blood flow affecting recovery. Its specific modulation in diseases presents potential therapeutic targets for drug development.

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.

9 product images

Western blot - Anti-PHD3 antibody [EPR17869] (ab184714)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/10000 dilution
Lane 1: PHD3 transfected HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate at 10 µg
Lane 2: Empty vector (vector control) transfected HEK-293 whole cell lysate at 10 µg
Lane 3: PHD3 transfected HEK-293 whole cell lysate treated with 0.1 mM CoCl2 for 4 hours at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 1s
Western blot - Anti-PHD3 antibody [EPR17869] (ab184714)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution
All lanes: Human fetal liver lysate at 10 µg
Secondary
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 15s
Western blot - Anti-PHD3 antibody [EPR17869] (ab184714)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/5000 dilution
All lanes: A549 (Human lung carcinoma) whole cell lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 2min
Western blot - Anti-PHD3 antibody [EPR17869] (ab184714)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution
Lane 1: Mouse pancreas lysate at 10 µg
Lane 2: Rat pancreas lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 1min
Western blot - Anti-PHD3 antibody [EPR17869] (ab184714)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution
Lane 1: Mouse kidney lysate at 10 µg
Lane 2: Mouse spleen lysate at 10 µg
Lane 3: Rat brain lysate at 10 µg
Lane 4: RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 5: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg
Lane 6: NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 15s
Western blot - Anti-PHD3 antibody [EPR17869] (ab184714)
Blocking/Dilution buffer: 5% NFDM/TBST.
PHD3 expression was induced by CoCl2 treatment (PMID: 18337469).
All lanes: Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution
Lane 1: Untreated MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg
Lane 2: MCF7 cell lysate treated with 0.5mM CoCl2 (Cobalt (II) chloride) for 6 hours at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 1min
Immunoprecipitation - Anti-PHD3 antibody [EPR17869] (ab184714)
PHD3 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with ab184714 at 1/70 dilution.
Western blot was performed from the immunoprecipitate using ab184714 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1/10000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10ug (Input).
Lane 2: ab184714 IP in NIH/3T3 whole cell lysate.
Lane 3: NIH/3T3 whole cell lysate supernatant after capture (unbound).
Lane 4: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab184714 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
ab184714 is not a strong binder for IP - only a partial amount of the target protein in the lysate was immune-precipitated.
All lanes: Immunoprecipitation - Anti-PHD3 antibody [EPR17869] (ab184714)
Predicted band size: 27 kDa
Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody [EPR17869] (ab184714)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma) cells labeling PHD3 with ab184714 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing weakly cytoplasm and nuclear staining on A549 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab184714 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody [EPR17869] (ab184714)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (Rat adrenal gland pheochromocytoma) cells labeling PHD3 with ab184714 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing weakly cytoplasm and nuclear staining on PC-12 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab184714 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.