HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody [JE63-25]
Catalog# HA722468
HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody [JE63-25]
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WB
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IHC-P
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IF-Cell
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IP
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FC
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Human
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unconjugated
Overview
Product Name
HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody [JE63-25]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human HIF Prolyl Hydroxylases aa 51-150 / 502.
Species Reactivity
Human
Validated Applications
WB, IHC-P, IF-Cell, IP, FC
Target Molecular Weight
Predicted band size: 47 kDa
Positive Control
HEK-293 cell lysate, A549 cell lysate, HCT 116 cell lysate, HeLa cell lysate, HEK-293, human lung cancer tissue, human colon cancer tissue.
Conjugation
unconjugated
Clone Number
JE63-25
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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
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IHC-P
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1:200
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IF-Cell
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1:100
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IP
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1-2μg/sample
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FC
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1:1,000
Target
Function
The product of this gene belongs to the family of prolyl 4-hydroxylases. This protein is a prolyl hydroxylase that may be involved in the degradation of hypoxia-inducible transcription factors under normoxia. It plays a role in adaptation to hypoxia and may be related to cellular oxygen sensing. Alternatively spliced variants encoding different isoforms have been identified. Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF1A at 'Pro-402' and 'Pro-564'. May function as a cellular oxygen sensor and, under normoxic conditions, may target HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. An autosomal recessive neurodevelopmental disorder characterized by global developmental delay, poor or absent speech, hypotonia, variable ocular movement and visual abnormalities, and respiratory difficulties. Disease onset is in infancy and death due to respiratory insufficiency may occur.
Background References
1. Koivunen P., Tiainen P., Hyvaerinen J., Williams K.E., Sormunen R., Klaus S.J., Kivirikko K.I., Myllyharju J. An endoplasmic reticulum transmembrane prolyl 4-hydroxylase is induced by hypoxia and acts on hypoxia-inducible factor alpha. J. Biol. Chem. 282:30544-30552 (2007)
Subcellular Location
Endoplasmic reticulum membrane.
UNIPROT
Synonyms
EGLN4 antibody
FLJ20262 antibody
HIF prolyl hydroxylase PH4 antibody
HIF-PH4 antibody
HIF-prolyl hydroxylase 4 antibody
HIFPH4 antibody
HPH-4 antibody
Hypoxia inducible factor prolyl 4 hydroxylase antibody
Hypoxia inducible factor prolyl hydroxylase antibody
Hypoxia-inducible factor prolyl hydroxylase 4 antibody
ExpandImages
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Western blot analysis of HIF Prolyl Hydroxylases on different lysates with Rabbit anti-HIF Prolyl Hydroxylases antibody (HA722468) at 1/1,000 dilution.
Lane 1: HEK-293 cell lysate
Lane 2: A549 cell lysate
Lane 3: HCT 116 cell lysate
Lane 4: HeLa cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: 25 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 (HA722468) 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. -
Immunocytochemistry analysis of HEK-293 cells labeling HIF Prolyl Hydroxylases with Rabbit anti-HIF Prolyl Hydroxylases antibody (HA722468) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HIF Prolyl Hydroxylases antibody (HA722468) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-HIF Prolyl Hydroxylases antibody (HA722468) 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 (HA722468) 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 human colon cancer tissue with Rabbit anti-HIF Prolyl Hydroxylases antibody (HA722468) 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 (HA722468) 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. -
HIF Prolyl Hydroxylases was immunoprecipitated from 0.2 mg HEK-293 cell lysate with HA722468 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722468 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HEK-293 cell lysate (input)
Lane 2: HA722468 IP in HEK-293 cell lysate
Lane 3: Rabbit IgG instead of HA722468 in HEK-293 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 26 seconds; ECL: K1801 -
Flow cytometric analysis of HEK-293 cells labeling HIF Prolyl Hydroxylases.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722468, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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