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Egl Nine Homolog 3 (C. Elegans) (EGLN3) (C-Term) antibody

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Synonyms zgc:77019, wu:fj78a08, EGLN3, phd3, PHD-3, PHD3, SM-20, 2610021G09Rik, AI505553, AI648162, Hif-p4h-3, Phd3, HIFP4H3, HIFPH3
(18), (14), (13), (5), (5), (3), (2), (2), (2), (1), (1)
(70), (27), (15), (5), (4), (2), (1), (1), (1)
(60), (16), (6)
(6), (5), (5), (3), (3), (3), (1)
Western Blotting (WB), Chromatin Immunoprecipitation (ChIP), Immunoprecipitation (IP)
(66), (46), (23), (5), (4), (2), (2), (1), (1)
Pubmed 7 references available
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Quantity 25 μL
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Immunogen A synthetic peptide made to the C-terminus of humanPHD3/HIF Prolyl Hydroxylase 3.
Cross-Reactivity (Details) Primate reactivity reported in scientific literature (PMID: 23732909). mmunogen sequence has 92% homology to rat.
Purification affinity purified
Alternative Name EGLN3 / PHD3 (EGLN3 Antibody Abstract)
Background HIF prolyl hydroxylase 3 is a prolyl hydroxylase that modifies HIF-alpha. Classic prolylhydroxylases are found in the endoplasmic reticulum and modify collagen, whereas HIFis an intracellular protein and the HPH sites do not resemble those modifying collagen. HIF is a transcriptional complex that plays a critical role in oxygen homeostasis. HPH isan essential component of the pathway through which cells sense oxygen. In thepresence of oxygen, HPHs convert specific prolyl residues in HIF-alpha tohydroxyproline, leading to HIF-alpha destruction. Low oxygen levels, sensed at thecellular level, cause the HIF conversion to be reduced so that HIF is stable and there isincreased angiogenesis.HIF prolyl hydroxylase 1, specifically, catalyzes the posttranslational formation of4-hydroxyproline in HIF alpha proteins. It hydroxylates HIF-1 alpha at Pro(564) andHIF-2 alpha. It targets HIF through the hydroxylation for proteasomal degradation viathe von Hippel-Lindau ubiquitylation complex. It may also play a role in cell growthregulation in muscle cells and in apoptosis in neuronal tissues, promoting cell deaththrough a caspase-dependent mechanism. Alternate Names: anti-EGL9 homolog of C. elegans 3 antibody, anti-EGLN3 antibody, anti-Factorresponsive smooth muscle protein antibody, anti-PHD3 antibody, anti-HIF ProlylHydroxylase 3 antibody, anti-HIFP4H3 antibody, anti-HIFPH3 antibody, anti-P4H3antibody, anti-Prolyl Hydroxylase Domain Containing Protein 3 antibody, anti-SM20antibody.
Gene Symbol: EGLN3
Gene ID 112399, 112407
Application Notes This PHD3/HIF Prolyl Hydroxylase 3 antibody is useful for Immunoprecipitation and Western blot, where a band is seen at ~27 kDa. This antibody has been tested against HeLa and MEF cell lysates. Immunoprecipitation was reported in PMID: 21575608. Chromatin Immunoprecipitation was reported in scientific literature.
Recommended dilutions: Chromatin Immunoprecipitation, Immunoprecipitation 1:10-1:500, Western Blot 1:500-1:2000
Protocol Protocol specific for HIF Prolyl Hydroxylase 3 Antibody Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 35 µg of sample lysate per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer Tris-citrate/Phosphate, pH 7-8, Sodium Azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Do not freeze.
Storage 4 °C
Supplier Images
Western Blotting (WB) image for anti-Egl Nine Homolog 3 (C. Elegans) (EGLN3) (C-Term) antibody (ABIN251418) Western Blot analysis of human PHD3, using ABIN151769. Samples: HeLa whole cell lysat...
Product cited in: Rodríguez-Jiménez, Moreno-Manzano, Lucas-Dominguez et al.: "Hypoxia causes downregulation of mismatch repair system and genomic instability in stem cells." in: Stem cells (Dayton, Ohio), Vol. 26, Issue 8, pp. 2052-62, 2008 (PubMed).

Mikhaylova, Ignacak, Barankiewicz et al.: "The von Hippel-Lindau tumor suppressor protein and Egl-9-Type proline hydroxylases regulate the large subunit of RNA polymerase II in response to oxidative stress." in: Molecular and cellular biology, Vol. 28, Issue 8, pp. 2701-17, 2008 (PubMed).

Minamishima, Moslehi, Bardeesy et al.: "Somatic inactivation of the PHD2 prolyl hydroxylase causes polycythemia and congestive heart failure." in: Blood, Vol. 111, Issue 6, pp. 3236-44, 2008 (PubMed).

Takeda, Aguila, Parikh et al.: "Regulation of adult erythropoiesis by prolyl hydroxylase domain proteins." in: Blood, Vol. 111, Issue 6, pp. 3229-35, 2008 (PubMed).

Ietta, Wu, Winter et al.: "Dynamic HIF1A regulation during human placental development." in: Biology of reproduction, Vol. 75, Issue 1, pp. 112-21, 2006 (PubMed).

Ivan, Haberberger, Gervasi et al.: "Biochemical purification and pharmacological inhibition of a mammalian prolyl hydroxylase acting on hypoxia-inducible factor." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, Issue 21, pp. 13459-64, 2002 (PubMed).

Background publications Bruick, McKnight: "A conserved family of prolyl-4-hydroxylases that modify HIF." in: Science (New York, N.Y.), Vol. 294, Issue 5545, pp. 1337-40, 2001 (PubMed).

Catalog No. ABIN251418
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