HIF1AN Recombinant Rabbit Monoclonal Antibody [JG37-66]
Catalog# ET7108-50
HIF1AN Recombinant Rabbit Monoclonal Antibody [JG37-66]
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WB
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IF-Cell
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IHC-P
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FC
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Human
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unconjugated
Overview
Product Name
HIF1AN Recombinant Rabbit Monoclonal Antibody [JG37-66]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human HIF1AN aa 1-50 / 349.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, FC
Molecular Weight
40 kDa
Positive Control
A549 cell lysate, Jurkat cell lysate, LOVO, SiHa, human prostate carcinoma tissue.
Conjugation
unconjugated
Clone Number
JG37-66
RRID
Product Features
Form
Liquid
Concentration
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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1:1,000-1:2,000
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IF-Cell
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1:50-1:200
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
Target
Function
Factor inhibiting HIF-1 (FIH-1) exists as a homodimer and binds to HIF-1α. Specifically, FIH-1 operates as an asparaginyl hydroxylase. It catalyzes the hydroxylation of the β-carbon of Asparagine residue 803 within the carboxy terminal transactivation domain of HIF-1α. This hydroxylation event blocks the association of HIF-1α with co-activators. FIH-1 also binds to von Hippel-Lindau (VHL) tumor suppressor protein, which represses transcriptional ac-tivity of HIF-1α. In transiently transfected human osteosarcoma cells, FIH-1 localizes to the cytoplasm. The structure of FIH-1 includes a jellyroll-like β-barrel containing ferrous-binding triad residues. The gene encoding human FIH-1 maps to chromosome 10q24.
Background References
1. Lando D et al. FIH-1 is an asparaginyl hydroxylase enzyme that regulates the transcriptional activity of hypoxia-inducible factor. Genes Dev 16:1466-1471 (2002).
2. Zheng X et al. Interaction with factor inhibiting HIF-1 defines an additional mode of cross-coupling between the Notch and hypoxia signaling pathways. Proc Natl Acad Sci USA 105:3368-3373 (2008).
Subcellular Location
Nucleus. Cytoplasm.
UNIPROT
Synonyms
DKFZp762F1811 antibody
Factor inhibiting HIF-1 antibody
Factor inhibiting HIF1 antibody
FIH 1 antibody
FIH-1 antibody
FIH1 antibody
FLJ20615 antibody
FLJ22027 antibody
HIF1AN antibody
HIF1N_HUMAN antibody
ExpandImages
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☑ Knockdown (KD)
Western blot analysis of HIF1AN on different lysates with Rabbit anti-HIF1AN antibody (ET7108-50) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-HIF1AN KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 40 kDa
Observed band size: 40 kDa
Exposure time: 40 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 (ET7108-50) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of HIF1AN on different lysates with Rabbit anti-HIF1AN antibody (ET7108-50) at 1/500 dilution.
Lane 1: A549 cell lysates
Lane 2: Jurkat cell lysates
Lysates/proteins at 10 µg/Lane.
Predicted band size: 40 kDa
Observed band size: 40 kDa
Exposure time: 2 minutes;
10% 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 (ET7108-50) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 5,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of LOVO cells labeling HIF1AN with Rabbit anti-HIF1AN antibody (ET7108-50) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-HIF1AN antibody (ET7108-50) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of SiHa cells labeling HIF1AN with Rabbit anti-HIF1AN antibody (ET7108-50) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-HIF1AN antibody (ET7108-50) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. -
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue with Rabbit anti-HIF1AN antibody (ET7108-50) at 1/50 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 (ET7108-50) at 1/50 dilution for 0.5 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"