Egl Nine Homolog 2 (C. Elegans) (EGLN2) (AA 300-400) antibody

Details for Product No. ABIN250637
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Antigen
Synonyms egln2, phd2, PHD1, 0610011A13Rik, C85656, Hif-p4h-1, Ier4, Phd1, SM-20, HIF-PH1, HPH-1, HPH-3, PHD-1, EIT6, HIFPH1
Epitope
AA 300-400
(14), (10), (10), (6), (5), (3), (2), (1), (1), (1), (1)
Reactivity
Mouse (Murine), Rat (Rattus)
(69), (28), (24), (14), (6), (2), (2), (1), (1)
Host
Rabbit
(68), (2), (1)
Clonality
Polyclonal
Conjugate
Un-conjugated
(5), (3), (3), (3), (3), (3), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
(60), (25), (11), (10), (10), (7), (3), (3), (2), (2)
Pubmed 1 reference available
Quantity 0.025 mL
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Catalog No. ABIN250637
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Immunogen A synthetic peptide made to an internal portion of mouse PHD2/HIF Prolyl Hydroxylase 2 (between residues 300-400).
Cross-Reactivity (Details) Predicted to react with rat,based on 100% sequence homology.
Purification affinity purified
Alternative Name EGLN2 / PHD1
Background HIF prolyl hydroxylase 2 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. HPH-2, specifically, catalyzes the posttranslational formation of 4-hydroxyproline in HIFalpha proteins. It hydroxylates HIF-1 alpha at Pro(402) and Pro(564), and HIF-2 alpha. Ittargets HIF through the hydroxylation for proteasomal degradation via the vonHippel-Lindau ubiquitylation complex. Alternate Names: anti-PHD2 antibody, anti-C1ORF12 antibody, anti-Chromosome 1 Open Reading Frame12 antibody, anti-EGL 9 homolog of C. elegans 1 antibody, anti-EGLN 1 EGLN1antibody, anti-HIF Prolyl Hydroxylase 2 antibody, anti-HIF-P4H 2 antibody, anti-HIFPH2antibody, anti-P4H2 antibody, anti-Prolyl Hydroxylase Domain Containing antibody,anti-PHD2 antibody, anti-EGLN1 antibody.
Gene Symbol: EGLN1
Gene ID 54583
UniProt Q91YE3
Application Notes This HIF Prolyl Hydroxylase 2 antibody is useful for Western blot, Immunocytochemistry/Immunofluorescence, Immunohistochemistry-Paraffin embedded sections and Immunoprecipitation. In Western blot a band is seen ~43 kDa representing HIF Prolyl Hydroxylase 2. There is also a non-specific band of similar intensity at ~75 kDa.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:50-1:500, Immunohistochemistry 2.5 µg/mL, Immunohistochemistry-Paraffin 2.5 µg/mL, Immunoprecipitation 1:10-1:500, Western Blot 2 µg/mL
Protocol Protocol specific for HIF Prolyl Hydroxylase 2 Antibody Western Blot Protocol
1. Perform SDS-PAGE (4-12 %) on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
. Rinse membrane with dH2O and then stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
. Rinse the blot in TBS for approximately 5 minutes.
. Block the membrane using 5 % non-fat dry milk + 1 % BSA in TBS for 2 hours at room temperature (RT).
. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.
. Dilute the rabbit anti-PHD2 (murine) primary antibody in blocking buffer and incubate 1 hour at RT.
. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.
. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at RT.
. Wash the blot in wash buffer [TBS + 0.1 % Tween] 3 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 manufacturer's instructions (we used BioFX Super Plus ECL). Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding. IHC-FFPE sectionsI. Deparaffinization: Western Blot Protocol
. Perform SDS-PAGE (4-12 %) on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
. Rinse membrane with dH2O and then stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
. Rinse the blot in TBS for approximately 5 minutes.
. Block the membrane using 5 % non-fat dry milk + 1 % BSA in TBS for 2 hours at room temperature (RT).
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
anti-Egl Nine Homolog 2 (C. Elegans) (EGLN2) (AA 300-400) antibody Immunohistochemistry: HIF Prolyl Hydroxylase 2 Antibody [ABIN250635SS] - Staining of renal tubular epithelium in mouse using ABIN250635 at 2.5ug/ml. Mouse kidney cortex, 40X.
anti-Egl Nine Homolog 2 (C. Elegans) (EGLN2) (AA 300-400) antibody (2) Western Blot: PHD2 Antibody [ABIN250635SS] - Detection of PHD2 in mouse kidney lysate using ABIN250635. ECL exposure, 20 seconds.
General 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).

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