抗EGLN3/PHD3抗体 | Anti-EGLN3/PHD3 Antibody

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	Western Blot: EGLN3/PHD3 Antibody [NB100-303] - Total protein from Mouse MEF, Human RT4 and HeLa, and Rat PC12 cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-PHD3 in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.
	Immunocytochemistry/Immunofluorescence: EGLN3/PHD3 Antibody [NB100-303] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-HIF Prolyl Hydroxylase 3 [NB100-303] at a 1:200 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
	Immunohistochemistry-Paraffin: EGLN3/PHD3 Antibody [NB100-303] - LDL Receptor was detected in immersion fixed paraffin-embedded sections of human liver cancer using rabbit anti-human antibody (Catalog # NB100-303) at 1:3000 dilution overnight at 4C. Tissue was stained using the VisuCyte anti-rabbit HRP polymer detection reagent (Catalog # VC003) with DAB chromogen (brown) and counterstained with hematoxylin (blue).Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.
	Western Blot: EGLN3/PHD3 Antibody [NB100-303] - Whole cell protein from MEF cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2 ug/ml anti-EGLN3/PHD3 in 1% milk, and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.
	Immunocytochemistry/Immunofluorescence: EGLN3/PHD3 Antibody [NB100-303] - analysis of PHD3 in ARPE-19 cells using anti-PHD3 antibody. Image from verified customer review.
	Simple Western: EGLN3/PHD3 Antibody [NB100-303] - Simple Western lane view shows a specific band for PHD3/HIF Prolyl Hydroxylase 3 in 0.5 mg/ml of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

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Background

HIF prolyl 4-hydroxylases (PHDs) are proyl hydroxylase domain-containing enzymes (PHD1/ Egln2, PHD2/ Egln1, PHD3/ Egln3, and P4H-TM) which are known for their role in mediating physiological responses to hypoxic stress via modulation of HIF1alpha expression levels. HIF-alpha subunit is regulated by hydroxylation, both by a family of PHDs leading to ubiquitination and proteasomal degradation, and by transcriptional inactivation following asparaginyl hydroxylation by FIH (factor inhibiting HIF). When oxygen levels are normal, HIF Prolyl Hydroxylase 3 (HIF-PH3/ PHD3/ Egln3) hydroxylates a specific proline found in HIF1A'a NODD/CODD domains and also hydroxylates HIF2A with preference for CODD site for HIF1A/HIF2A. Once hydroxylated, HIFs undergo proteasomal degradation via von Hippel-Lindau (VHL) ubiquitination complex. Upon hypoxic trigger, the hydroxylation reaction is tempered which let HIFs to escape degradation process leading to their nuclear translocation, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. HIF-PH3 is the most important isozyme in limiting physiological activation of HIFs (particularly HIF2A) in hypoxia. Besides HIFs, in hypoxic conditions HIF-PH3 also hydroxylates PKM (thereby limiting glycolysis) and hydroxylates/regulates the stability of ADRB2. In cardiomyocytes, HIF-PH3 inhibits Bcl2's anti-apoptotic effect by disrupting the BAX-BCL2 complex, and in neurons, HIF-PH3 has a NGF-induced proapoptotic effect mediated via CASP3 activity regulation. HIF-PH3 also plays a crucial role in DNA damage response by hydroxylating TELO2, promoting its interaction with ATR which is required for activation of the ATR/CHK1/p53 pathway.

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