Key features and details
- Mouse monoclonal [mAbcam74796] to PHD2 / prolyl hydroxylase
- Suitable for: WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Product nameAnti-PHD2 / prolyl hydroxylase antibody [mAbcam74796]
See all PHD2 / prolyl hydroxylase primary antibodies
DescriptionMouse monoclonal [mAbcam74796] to PHD2 / prolyl hydroxylase
Tested applicationsSuitable for: WBmore details
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human PHD2/ prolyl hydroxylase aa 400 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available as
- This antibody gave a positive signal in human skeletal muscle tissue lysate, and in the following whole cell lysates: HepG2; HeLa; U20S.
This antibody clone is manufactured by Abcam.
This monoclonal antibody to PHD2 has been knockout validated in Western blot. The expected band for PHD2 was observed in wild type cells and the band was not seen in knockout cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
Concentration information loading...
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab74796 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 5 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 46 kDa).|
FunctionCatalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF-1 alpha at 'Pro-402' and 'Pro-564', and HIF-2 alpha. Functions as a cellular oxygen sensor and, under normoxic conditions, targets HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitination complex.
Tissue specificityAccording to PubMed:11056053, widely expressed with highest levels in skeletal muscle and heart, moderate levels in pancreas, brain (dopaminergic neurons of adult and fetal substantia nigra) and kidney, and lower levels in lung and liver. According to PubMed:12351678 widely expressed with highest levels in brain, kidney and adrenal gland. Expressed in cardiac myocytes, aortic endothelial cells and coronary artery smooth muscle.
Involvement in diseaseDefects in EGLN1 are the cause of erythrocytosis familial type 3 (ECYT3) [MIM:609820]. ECYT3 is an autosomal dominant disorder characterized by increased serum red blood cell mass, elevated serum hemoglobin and hematocrit, and normal serum erythropoietin levels.
Sequence similaritiesContains 1 Fe2OG dioxygenase domain.
Contains 1 MYND-type zinc finger.
- Information by UniProt
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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 - ab74796 observed at 52 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab74796 detected the expected band for PHD2 in wild type cells and the band was not seen in knockout cells. Additional cross reactive bands were observed in the wild type and knockout cells. Wild-type and PHD2 / prolyl hydroxylase knockout samples were subjected to SDS-PAGE. ab74796 at a concentration of 5 µg/ml and 181602 (loading control to GAPDH) diluted to 1/10,000 were incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-PHD2 / prolyl hydroxylase antibody [mAbcam74796] (ab74796) at 5 µg/ml
Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330)
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 44,52 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
The 52-kDa band observed is comparable to the band detected by other commercially available antibodies to human prolyl hydroxylase (PHD2). Based on the immunogen sequence, this antibody is predicted to cross react with both isoform 1 (46 kDa) and isoform 2 (44 kDa) of PHD2. We believe that the band observed around 44-kDa corresponds to isoform 2.
ab74796 has not yet been referenced specifically in any publications.