PHD2 Antibody - #DF6285

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製品: PHD2 Antibody
カタログ: DF6285
タンパク質の説明: Rabbit polyclonal antibody to PHD2
アプリケーション: WB IHC IF/ICC
反応性: Human, Mouse, Rat
予測: Pig, Bovine, Horse, Sheep, Rabbit, Dog
分子量: 46kDa; 46kD(Calculated).
ユニプロット: Q9GZT9
RRID: AB_2838251

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MS Report
HPLC Report
MSDS
説明書
COA
プロトコル
WB Handbook
IHC Protocol
IF/ICC Protocol

製品説明

ソース:
Rabbit
アプリケーション:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat
予測:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%)
クローナリティ:
Polyclonal
特異性:
PHD2 Antibody detects endogenous levels of total PHD2.
RRID:
AB_2838251
引用形式: Affinity Biosciences Cat# DF6285, RRID:AB_2838251.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ／展開

C1ORF12; Chromosome 1 Open Reading Frame 12; DKFZp761F179; ECYT 3; ECYT3; EGL 9 homolog of C. elegans 1; EGL nine (C.elegans) homolog 1; Egl nine homolog 1 (C. elegans); Egl nine homolog 1; Egl nine like protein 1; EGLN 1; Egln1; EGLN1_HUMAN; HIF PH2; HIF Prolyl Hydroxylase 2; HIF-PH2; HIF-prolyl hydroxylase 2; HIFP4H 2; HIFPH2; HPH 2; HPH-2; HPH2; Hypoxia inducible factor prolyl hydroxylase 2; Hypoxia-inducible factor prolyl hydroxylase 2; ORF13; P4H2; PHD 2; PhD2; PNAS 118; PNAS 137; Prolyl Hydroxylase Domain Containing Protein 2; Prolyl hydroxylase domain-containing protein 2; Rat Homolog of SM20; SM 20; SM-20; SM20; Zinc finger MYND domain containing protein 6; ZMYND6;

免疫原

免疫原:
Uniprot:

Q9GZT9(EGLN1_HUMAN) >>Visit HPA database.

遺伝子(ID):
EGLN1(54583)
発現特異性:
Q9GZT9 EGLN1_HUMAN:

According 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. According to PubMed:12788921; expressed in adult and fetal heart, brain, liver, lung, skeletal muscle and kidney. Also expressed in placenta. Highest levels in adult heart, brain, lung and liver and fetal brain, heart spleen and skeletal muscle.

タンパク質の説明:
PHD1 (Egln2), PHD-2 (Egln1), and PHD3 (Egln3) are members of the Egln family of proline hydroxylases. They function as oxygen sensors that catalyze the hydroxylation of HIF on prolines 564 and 402, initiating the first step of HIF degradation through the VHL/ubiquitin pathway (1,2). PHD1 is highly expressed in a wide array of tissues whereas PHD2 and PHD3 are expressed mainly in heart and skeletal muscle (1,3). The mRNA levels of PHD are upregulated by HIF through the hypoxia-response element under low oxygen conditions (4-7). These three enzymes also exhibit different peptide specificity target proteins, PHD1 and PHD2 can hydroxylate both proline 402 and proline 564, but PHD3 can only hydroxylate proline 564 (2,8). In addition to HIF, PHD enzymes have also has been shown to catalyze the hydroxylation of RNA polymerase subunits and myogenin (3,9).
タンパク質配列:
MANDSGGPGGPSPSERDRQYCELCGKMENLLRCSRCRSSFYCCKEHQRQDWKKHKLVCQGSEGALGHGVGPHQHSGPAPPAAVPPPRAGAREPRKAAARRDNASGDAAKGKVKAKPPADPAAAASPCRAAAGGQGSAVAAEAEPGKEEPPARSSLFQEKANLYPPSNTPGDALSPGGGLRPNGQTKPLPALKLALEYIVPCMNKHGICVVDDFLGKETGQQIGDEVRALHDTGKFTDGQLVSQKSDSSKDIRGDKITWIEGKEPGCETIGLLMSSMDDLIRHCNGKLGSYKINGRTKAMVACYPGNGTGYVRHVDNPNGDGRCVTCIYYLNKDWDAKVSGGILRIFPEGKAQFADIEPKFDRLLFFWSDRRNPHEVQPAYATRYAITVWYFDADERARAKVKYLTGEKGVRVELNKPSDSVGKDVF

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Xenopus
67
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9GZT9 基板として

Site PTM Type Enzyme
A2 Acetylation
S5 Phosphorylation
S12 Phosphorylation
S14 Phosphorylation
S61 Phosphorylation
S125 Phosphorylation
C127 S-Nitrosylation
S174 Phosphorylation
K216 Ubiquitination
T232 Phosphorylation
K234 Ubiquitination
K244 Methylation
K244 Ubiquitination
K255 Ubiquitination
K286 Ubiquitination
T308 Phosphorylation
K337 Ubiquitination
K350 Ubiquitination
K402 Ubiquitination
T405 Phosphorylation
K408 Acetylation
K408 Ubiquitination
K416 Ubiquitination

研究背景

機能:

Cellular oxygen sensor that catalyzes, under normoxic conditions, the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A. Also hydroxylates HIF2A. Has a preference for the CODD site for both HIF1A and HIF1B. Hydroxylated HIFs are then targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. Under hypoxic conditions, the hydroxylation reaction is attenuated allowing HIFs to escape degradation resulting in their translocation to the nucleus, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. EGLN1 is the most important isozyme under normoxia and, through regulating the stability of HIF1, involved in various hypoxia-influenced processes such as angiogenesis in retinal and cardiac functionality. Target proteins are preferentially recognized via a LXXLAP motif.

PTMs:

S-nitrosylation inhibits the enzyme activity up to 60% under aerobic conditions. Chelation of Fe(2+) has no effect on the S-nitrosylation. It is uncertain whether nitrosylation occurs on Cys-323 or Cys-326.

細胞の位置付け:

Cytoplasm. Nucleus.
Note: Mainly cytoplasmic. Shuttles between the nucleus and cytoplasm (PubMed:19631610). Nuclear export requires functional XPO1.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

According to 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 towidely expressed with highest levels in brain, kidney and adrenal gland. Expressed in cardiac myocytes, aortic endothelial cells and coronary artery smooth muscle. According to; expressed in adult and fetal heart, brain, liver, lung, skeletal muscle and kidney. Also expressed in placenta. Highest levels in adult heart, brain, lung and liver and fetal brain, heart spleen and skeletal muscle.

サブユニット構造:

Monomer. Interacts with ING4; the interaction inhibits the hydroxylation of HIF alpha proteins. Interacts with PTGES3 (via PXLE motif); thereby recruiting EGLN1 to the HSP90 pathway to facilitate HIF alpha proteins hydroxylation. Interacts with LIMD1. Found in a complex composed of LIMD1, VHL, EGLN1/PHD2, ELOB and CUL2. Interacts with EPAS1. Interacts with CBFA2T3. Interacts with HIF1A.

タンパク質ファミリー:

The beta(2)beta(3) 'finger-like' loop domain is important for substrate (HIFs' CODD/NODD) selectivity.

研究領域

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Renal cell carcinoma.   (View pathway)

参考文献

1). ETV2 regulating PHD2-HIF-1α axis controls metabolism reprogramming promotes vascularized bone regeneration. Bioactive Materials, 2024 [IF=18.9]

Application: WB    Species: Mouse    Sample:

Fig. 2. ETV2 induces HIF-1α stabilization and nuclear accumulation by transcriptional inhibition of PHD2 and ERK1/2 phosphorylation (A) KEGG functional enrichment analysis of RNA sequence (B) A heatmap displays genes associated with osteogenesis and HIF-1 signaling (C, D) The protein and mRNA expression of HIF-1α and PHDs following ETV2 overexpression (E) Schematic of putative ETV2 binding elements on PHD2 promoter region (F) Dual luciferase reporter gene assay of ETV2 and PHD2 (G) Cytoplasmic HIF-1α, total and phosphorylated ERK1/2, and intracellular HIF-1α expression post ETV2 overexpression (H) Representative immunofluorescent images of intracellular phosphorylated ERK1/2. The Cy3 channel fluorescence represents lentiviral transfection. Scale bar: 50 μm (I) Representative immunofluorescent images of intranuclear HIF-1α. The Cy3 channel fluorescence represents lentiviral transfection. The orange arrows indicate the fluorescence of HIF-1α in the nucleus, while the white arrows point to the absence of HIF-1α fluorescence in the nucleus. Scale bar: 50 μm (NS, no significant difference, NC, negative control; OE, overexpression. Data are presented as the mean of >3 independent experiments ±SD. *P < 0.05, **P < 0.01, and ***P < 0.001).

Application: IF/ICC    Species: Mouse    Sample:

Fig. 2. ETV2 induces HIF-1α stabilization and nuclear accumulation by transcriptional inhibition of PHD2 and ERK1/2 phosphorylation (A) KEGG functional enrichment analysis of RNA sequence (B) A heatmap displays genes associated with osteogenesis and HIF-1 signaling (C, D) The protein and mRNA expression of HIF-1α and PHDs following ETV2 overexpression (E) Schematic of putative ETV2 binding elements on PHD2 promoter region (F) Dual luciferase reporter gene assay of ETV2 and PHD2 (G) Cytoplasmic HIF-1α, total and phosphorylated ERK1/2, and intracellular HIF-1α expression post ETV2 overexpression (H) Representative immunofluorescent images of intracellular phosphorylated ERK1/2. The Cy3 channel fluorescence represents lentiviral transfection. Scale bar: 50 μm (I) Representative immunofluorescent images of intranuclear HIF-1α. The Cy3 channel fluorescence represents lentiviral transfection. The orange arrows indicate the fluorescence of HIF-1α in the nucleus, while the white arrows point to the absence of HIF-1α fluorescence in the nucleus. Scale bar: 50 μm (NS, no significant difference, NC, negative control; OE, overexpression. Data are presented as the mean of >3 independent experiments ±SD. *P < 0.05, **P < 0.01, and ***P < 0.001).
2). Application of Multiparametric Magnetic Resonance Imaging to Monitor the Early Antitumor Effect of CuS@GOD Nanoparticles in a 4 T1 Breast Cancer Xenograft Model. JOURNAL OF MAGNETIC RESONANCE IMAGING, 2022 (PubMed: 34427359) [IF=4.4]

Application: IHC    Species: Mouse    Sample: 4 T1 breast cancer cells

FIGURE 4: The histological examination at the time points corresponding to the MRI scan. The histologically stained (?200) sections of H&E, Ki67 and TUNEL at 24 hours, VEGF at 1 hour, and EGLN1 at 0 hour after treatment (a). The positive staining rate (%) of Ki67 (b) and TUNEL (c) and the mean IOD of VEGF (d) and EGLN1 (e) staining of the four experimental groups at different time points. * indicates a significant difference between groups at the same time point. Group 1: normal saline, group 2:CuS@GOD NPs, group 3: CuS NPs + laser and group 4: CuS@GOD NPs + laser.
3). Downregulation of Proline Hydroxylase 2 and Upregulation of Hypoxia-Inducible Factor 1α are Associated with Endometrial Cancer Aggressiveness. Cancer Management and Research, 2019 (PubMed: 31819628) [IF=3.3]

Application: WB    Species: human    Sample: endometrial cancer

Figure 2| Western blot analysis of PHD2 and HIF-1α protein expression in normal endometrium, atypical endometrial hyperplasia, and endometrial cancer. (A) PHD2 expression was reduced in endometrial cancer compared with normal endometrium (p<0.05, chi-square test).

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