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Anti-EGLN1 Antibody EPR3661
Also for EGLN1 (NM_022051)
|A synthetic peptide corresponding to residues on the C-terminus in human PHD2 was used as an immunogen.|
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||ICC/IF: 1:200; WB: 1:1000 - 1:10000; IP: 1:10 - 1:100; IHC-P: 1:50 - 1:100; ICC: 1:100 - 1:250; FC: 1:100
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Does not react with Rat
|Homo sapiens egl-9 family hypoxia-inducible factor 1 (EGLN1)|
|C1orf12; ECYT3; HIF-PH2; HIFPH2; HPH-2; HPH2; PHD2; SM20; ZMYND6|
|PHD2 (Egl nine homolog 1) catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. HIF is a transcriptional complex that plays a central role in mammalian oxygen homeostasis. This protein functions as a cellular oxygen sensor, and under normal oxygen concentration, modification by prolyl hydroxylation is a key regulatory event that targets HIF subunits for proteasomal destruction via the von Hippel-Lindau ubiquitylation complex. Mutations in PHD2 are associated with erythrocytosis familial type 3 (ECYT3).|
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Western blot - PHD2 / prolyl hydroxylase antibody [EPR3661]; All lanes : Anti-PHD2 / prolyl hydroxylase antibody [EPR3661] at 1/1000 dilution.Lane 1 : SH-SY5Y cell lysate, treated with CoCl2.Lane 2 : SH-SY5Y cell lysate.Lane 3 : A549 cell lysate.Lysates/proteins at 10 µg per lane.Predicted band size : 46 kDa.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PHD2 / prolyl hydroxylase antibody [EPR3661]; ab109088 at 1/50 dilution staining PHD2 / prolyl hydroxylase in Human kidney by Immunohistochemistry, Paraffin-embedded tissue
Immunocytochemistry/ Immunofluorescence - Anti-PHD2 / prolyl hydroxylase antibody [EPR3661]; ICC/IF image of ab109088 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab109088 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight? 488 goat anti- rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43uM.
Flow Cytometry - Anti-PHD2 / prolyl hydroxylase antibody [EPR3661]; Overlay histogram showing SH-SY5Y cells stained with ab109088 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.