PHD1 / prolyl hydroxylase Recombinant Rabbit Monoclonal Antibody [SP00-48]
Catalog# ET1604-5
PHD1 / prolyl hydroxylase Recombinant Rabbit Monoclonal Antibody [SP00-48]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
PHD1 / prolyl hydroxylase Recombinant Rabbit Monoclonal Antibody [SP00-48]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human PHD1 aa 1-91 / 407.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC
Molecular Weight
Predicted band size: 44 kDa
Positive Control
HeLa cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Mouse heart tissue lysate, Rat brain tissue lysate, Rat heart tissue lysate, Hela, A549, SKOV-3, human breast cancer tissue, mouse testis tissue.
Conjugation
unconjugated
Clone Number
SP00-48
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:5,000
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IF-Cell
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1:50-1:200
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IF-Tissue
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1:50-1:200
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IHC-P
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1:1,000
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FC
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1:50-1:100
Target
Function
Prolyl hydroxylase domain proteins HIF PHD1, HIF PHD2 and HIF PHD3 (known as PHD1, PHD2 and PHD3 in rodents, respectively) can hydroxylate HIF-α subunits. Hypoxia-inducible factor (HIF) is a transcriptional regulator important in several aspects of oxygen homeostasis. The prolyl hydroxylases catalyze the posttranslational formation of 4-hydroxyproline in HIF-α proteins. HIF PHD1, which is widely expressed, with highest levels of expression in testis, functions as a cellular oxygen sensor and is important in cell growth regulation. HIF PHD1 can localize to the nucleus or the cytoplasm and is also detected in hormone responsive tissues, such as normal and cancerous mammary, ovarian and prostate epithelium. HIF PHD1 is encoded by EGLN2, which maps to chromosome 19q13.3. HIF PHD2 is regarded as the main cellular oxygen sensor, as RNA interference against HIF PHD2, but not HIF PHD1 or HIF PHD3, is enough to stabilize HIF-1α in normoxia. HIF PHD2, a direct HIF target gene, is expressed mainly in skeletal muscle, heart, kidney and brain. HIF PHD3 may play a role in the regulation of cell growth in muscle cells and in apoptosis in neuronal tissue. HIF PHD3 is widely expressed, although the highest levels can be detected in placenta and he
Background References
1. Lalevée S et al. miR455 is linked to hypoxia signaling and is deregulated in preeclampsia. Cell Death Dis 5:e1408 (2014).
2. Van Welden S et al. Differential expression of prolyl hydroxylase 1 in patients with ulcerative colitis versus patients with Crohn\'s disease/infectious colitis and healthy controls. J Inflamm (Lond) 10:36 (2013).
Tissue Specificity
Expressed in adult and fetal heart, brain, liver, lung, skeletal muscle, and kidney. Also expressed in testis and placenta. Highest levels in adult brain, placenta, lung, kidney, and testis. Expressed in hormone responsive tissues, including normal and cancerous mammary, ovarian and prostate epithelium.
Post-translational Modification
Ubiquitinated by SIAH1 and/or SIAH2 in response to the unfolded protein response (UPR), leading to its degradation.
Subcellular Location
Nucleus.
Synonyms
DKFZp434E026 antibody
EGL nine (C.elegans) homolog 2 antibody
Egl nine homolog 2 (C. elegans) antibody
Egl nine homolog 2 antibody
EGLN 2 antibody
EGLN2 antibody
EGLN2_HUMAN antibody
EIT 6 antibody
EIT6 antibody
Estrogen-induced tag 6 antibody
ExpandImages
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Western blot analysis of PHD1 / prolyl hydroxylase on different lysates with Rabbit anti-PHD1 / prolyl hydroxylase antibody (ET1604-5) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
Lane 3: PC-12 cell lysate (20 µg/Lane)
Lane 4: Mouse brain tissue lysate (40 µg/Lane)
Lane 5: Mouse heart tissue lysate (40 µg/Lane)
Lane 6: Rat brain tissue lysate (40 µg/Lane)
Lane 7: Rat heart tissue lysate (40 µg/Lane)
Predicted band size: 44 kDa
Observed band size: 50 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1604-5) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
ICC staining of PHD1 / prolyl hydroxylase in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1604-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of PHD1 / prolyl hydroxylase in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1604-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of PHD1 / prolyl hydroxylase in SKOV-3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1604-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-PHD1 / prolyl hydroxylase antibody (ET1604-5) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-5) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PHD1 / prolyl hydroxylase antibody (ET1604-5) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-5) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of PHD1 / prolyl hydroxylase was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1604-5, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"