FIH-1/HIF-1AN Antibody

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Analysis of astrocytes lysate from Rat brain using FIH-1/HIF-1AN antibody at 1:500 dilution with ECL detection. The antibody detected a major band at ~40 kDa position representing FIH-1/HIF-1AN protein.

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Representative picture of western blot in histopathologically unchanged tissue (N) and primary cancerous tissue (C) from patients with CRC. Immunodetection of bands was performed with Rp anti- PHD1 (Catalog # NB100-310), - PHD2 (Catalog # NB100-137), - PHD3 (Catalog # NB100-139) and -FIH-1/HIF-1AN Ab (Catalog # NB100-139), followed by incubation with goat anti-rabbit HRP-conjugated Ab. The membrane was stripped and incubated with Rp anti-GAPDH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. Image collected and cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-13-526), licensed under a CC-BY licence.

Immunohistochemistry: FIH-1/HIF-1AN Antibody [NB100-428] - Detection of FIH-1 in rat brain tissue.

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Rat VSMC (vascular smoot muscle) cells treated (siFIH) or not (CTL) with a specific siRNA against FIH-1. Western blot image submitted by a verified customer review.

• Reactivity: Hu, Mu, Rt
• Applications: WB, EM, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

FIH-1/HIF-1AN Antibody Summary

• Immunogen: A synthetic peptide made to the C-terminal region of human Factor Inhibiting HIF-1 (between residues 300 and the C-terminus). [UniProt# Q9NWT6]
• Localization: Nuclear
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: HIF1AN
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Electron Microscopy
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Frozen 1:50-1:200
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Immunoprecipitation 1:10-1:500
  • Knockout Validated
  • Western Blot 1:1000-1:5000
• Application Notes: Knockout data from PMID 31937588.
• Theoretical MW: 40.3 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  FIH-1/HIF-1AN Overexpression Lysate
  HIF-1 alpha Knockout CoCl2-treated/untreated HeLa Cell Lysate
  HIF-1 alpha Knockout Hypoxic-treated/untreated HeLa Cell Lysate
  NIH 3T3 CoCl2 treated/Untreated Cell Lysate
  L929 CoCl2 treated/Untreated Cell Lysate
  Raw 264.7 CoCl2 treated/Untreated Cell Lysate
  PC12 CoCl2 treated/Untreated Cell Lysate
  PC12 Hypoxic / Normoxic Cell Lysate
  Raw 264.7 Hypoxic / Normoxic Cell Lysate
  L929 Hypoxic / Normoxic Cell Lysate
  NIH 3T3 Hypoxic / Normoxic Cell Lysate

• Control Peptide:

  FIH-1/HIF-1AN Peptide (NB100-428PEP)

Reactivity Notes

Human, rat and mouse (PMID 19720742).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Tris-Citrate/Phosphate (pH 7.0 - 8.0)
• Preservative: 0.1% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for FIH-1/HIF-1AN Antibody

• DKFZp762F1811
• EC 1.14.11.16
• factor inhibiting HIF1
• Factor inhibiting HIF-1
• FIH1
• FIH-1
• FIH1FIH-1
• FLJ20615
• FLJ22027
• HIF1AN
• HIF-1AN
• hypoxia inducible factor 1, alpha subunit inhibitor
• hypoxia-inducible factor 1-alpha inhibitor
• Hypoxia-inducible factor asparagine hydroxylase
• Peptide-aspartate beta-dioxygenase

Background

Factor inhibiting HIF1 (FIH1) is an asparaginyl hydroxylase enzyme that regulates HIF's transcriptional activity by hydroxylating HIF-1 alpha at 'Asp-803' in the CAD (C-terminal transactivation domain). Hydroxylation via PHDs followed by proteosome degradation is another mechanism that regulates HIF1, however, FIH1 is able to exercise the control even under severe hypoxic conditions, when PHD enzymes fail to do so. Localized mainly in cytoplasm, FIH1 functions as oxygen sensor and FIH1 interaction with NOTCH1 results in its nuclear localization whereas FIH1-ABPA3 interaction leads to its perinuclear localization. During normoxic conditions, FIH1 hydroxylates a conserved asparaginyl residue within CTAD of HIF-1 alpha/HIF-2 alpha, leading to steric clash that prevents the recruitment of the co-activators p300 and CBP. On the other hand, under hypoxic conditions, FIH1 is inactive, which results in activation of HIF-alpha signaling. FIH1 is involved in transcriptional repression through interaction with HIF1A, VHL and HDAC and hydroxylates specific Asn residues within ARD (ankyrin repeat domains) of NFKB1, NFKBIA, NOTCH1, ASB4, PPP1R12A as well as other ARD-containing proteins. FIH1 also hydroxylates Asp and His residues within ARDs of ANK1 and TNKS2, respectively. FIH1 has the ability to negatively regulate NOTCH1 activity thus accelerating myogenic differentiation, and positively regulates ASB4 activity which promotes vascular differentiation. FIH1-Mint3 binding affects FIH1's ability to modify HIF-1 alpha in an oxygen-independent manner and FIH1-Bax interaction plays inhibitory role in apoptosis.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for FIH-1/HIF-1AN Antibody (NB100-428)(25)

Publications using NB100-428

• Li ZL, Wang B, Lv LL et al. FIH-1-modulated HIF-1 alpha C-TAD promotes acute kidney injury to chronic kidney disease progression via regulating KLF5 signaling Acta pharmacologica Sinica Mar 3 2021 [PMID: 33658705]
• Chung Y, Saitoh Y, Hayashi T et al. Mint3 is dispensable for pancreatic and kidney functions in mice Biochemistry and biophysics reports Dec 1 2020 [PMID: 33319072] (WB)
• Mader J, Huber J, Bonn F et al. Oxygen-dependent asparagine hydroxylation of the ubiquitin-associated (UBA) domain in Cezanne regulates ubiquitin binding J. Biol. Chem. Jan 14 2020 [PMID: 31937588] (WB, KO, Human)
• Docherty C K, Nilsen M et al. Influence of 2-Methoxyestradiol and Sex on Hypoxia-Induced Pulmonary Hypertension and Hypoxia-Inducible Factor-1-alpha. J Am Heart Assoc May 3 2019 [PMID: 30819028] (WB, Human)
• Pillai MR, Mihi B, Ishiwata K et al. Myc-induced nuclear antigen constrains a latent intestinal epithelial cell-intrinsic anthelmintic pathway PLoS ONE Feb 26 2019 [PMID: 30807587] (WB, Mouse)
• Bullen JW, Tchernyshyov I, Holewinski RJ et al. Protein kinase A-dependent phosphorylation stimulates the transcriptional activity of hypoxia-inducible factor 1 Sci Signal 2016 May 31 [PMID: 27245613] (WB, Human)
• Nakaoka HJ, Hara T, Yoshino S et al. NECAB3 Promotes Activation of Hypoxia-inducible factor-1 during Normoxia and Enhances Tumourigenicity of Cancer Cells Sci Rep Mar 7 2016 [PMID: 26948053] (IP, Human)
• Kiriakidis S, Henze AT, Kruszynska-Ziaja I et al. Factor-inhibiting HIF-1 (FIH-1) is required for human vascular endothelial cell survival FASEB J. 2015 Apr 02 [PMID: 25837583] (WB, Human)
• Karttunen S, Duffield M, Scrimgeour Nr et al. Oxygen-dependent hydroxylation by Factor Inhibiting HIF (FIH) regulates the TRPV3 ion channel. J. Cell. Sci. 2014 Nov 20 [PMID: 25413349] (WB)
• Rawluszko AA, Bujnicka KE, Horbacka K et al. Expression and DNA methylation levels of prolyl hydroxylases PHD1, PHD2, PHD3 and asparaginyl hydroxylase FIH in colorectal cancer. BMC Cancer. 2013 Nov 6 [PMID: 24195777] (WB, Human)
• Janke K, Brockmeier U, Kuhlmann K et al. Factor Inhibiting HIF-1 (FIH-1) modulates protein interactions of Apoptosis-Stimulating p53 binding Protein 2 (ASPP2). J Cell Sci 2013 Apr 19 [PMID: 23606740] (WB, Human)
• Nishida C, Kusubata K, Tashiro Y et al. MT1-MMP plays a critical role in hematopoiesis by regulating HIF-mediated chemokine/cytokine gene transcription within niche cells Blood 2012 Jun 7 [PMID: 22544701]
• Lim CS, Kiriakidis S, Paleolog EM, Davies AH. Increased activation of the hypoxia-inducible factor pathway in varicose veins. J Vasc Surg;55(5):1427-1439e1. 2012 May. [PMID: 22277691] (WB, Human)
• Wilkins SE, Karttunen S, Hampton-Smith RJ et al. Factor inhibiting HIF (FIH) recognises distinct molecular features within hypoxia inducible factor (HIF)-[alpha] versus ankyrin repeat substrates. The Journal of Biological Chemistry. 2012 Jan 23. [PMID: 22270367]
• Melvin A, Mudie S, Rocha S. The chromatin remodeler ISWI regulates the cellular response to hypoxia: role of FIH. Mol Biol Cell 2011 Nov [PMID: 21900490] (Human)
• Singleton RS, Trudgian DC, Fischer R et al. Quantitative mass spectrometry reveals the dynamics of Factor Inhibiting HIF (FIH)-catalysed hydroxylation. J Biol Chem. 2011 Jul 30. [PMID: 21808058]
• Schodel J, Bohr D, Klanke B et al. Factor inhibiting HIF limits the expression of hypoxia-inducible genes in podocytes and distal tubular cells. Kidney Int. 2010 [PMID: 20720525]
• Luo T, Zhang X, Nicholson WL et al. Molecular characterization of antibody epitopes of Ehrlichia chaffeensis ankyrin protein 200 and tandem repeat protein 47 and evaluation of synthetic immunodeterminants for serodiagnosis of human monocytotropic ehrlichiosis. Clin Vaccine Immunol 2010 Jan [PMID: 19955322]
• Minamishima YA, Moslehi J, Padera RF et al. A feedback loop involving the Phd3 prolyl hydroxylase tunes the mammalian hypoxic response in vivo. Mol Cell Biol;29(21):5729-41. 2009 Nov. [PMID: 19720742] (WB, Mouse)
• Chan DA, Kawahara TL, Sutphin PD et al. Tumor vasculature is regulated by PHD2-mediated angiogenesis and bone marrow-derived cell recruitment. Cancer Cell 2009 Jun 2 [PMID: 19477431]
• Xu X, Vaithiyalingam S, Glick GG, Mordes DA, Chazin WJ, Cortez D. The basic cleft of RPA70N binds multiple checkpoint proteins, including RAD9, to regulate ATR signaling. Mol Cell Biol;28(24):7345-53. 2008 Dec. [PMID: 18936170]
• Zheng X, Linke S, Dias JM 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 U S A;105(9):3368-73. 2008 Mar 4. [PMID: 18299578] (WB)
• Li, J et al. Protein kinase C-mediated modulation of FIH-1 expression by the homeodomain protein CDP/Cut/Cux. Mol Cell Biol;27(20):7345-53. 2007 Oct. [PMID: 17682059]
• Ferguson III, JE, Wu, Y et al. ASB4 is a hydroxylation substrate of FIH and promotes vascular differentiation via an oxygen-dependent mechanism. Mol Cell Biol;27(18):6407-19. 2007 Sep. [PMID: 17636018]
• Khan Z, Michalopoulos GK, Stolz DB. Peroxisomal localization of hypoxia-inducible factors and hypoxia-inducible factor regulatory hydroxylases in primary rat hepatocytes exposed to hypoxia-reoxygenation. Am J Pathol. 2006 Oct [PMID: 17003483] (WB, IHC-P, ICC/IF, EM, Rat)

Reviews for FIH-1/HIF-1AN Antibody (NB100-428) (3)

Average Rating: 4.3

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Rat.

reviewed by: Darren Richard WB Rat 06/05/2020

Summary

• Application: Western Blot
• Sample Tested: Vascular Smooth Muscle Cells,Whole cell lysate
• Species: Rat
• Lot: A3

reviewed by: Verified Customer WB Human 11/12/2017

Summary

• Application: Western Blot
• Sample Tested: Cell lysates from prostate cancer cell lines
• Species: Human
• Lot: A2

Comments

• Comments: Sample: Whole cell lysates of prostate cancer cell lines (20ug)
  Blocking: 5% nonfat dry milk reconstituted in TBS-T for 1 hour at room temperature
  Primary Antibody diluted (1:500) in 5% nonfat dry milk reconstituted in TBS-T overnight at 4deg C
  Secondary Antibody: Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution in TBS-T

reviewed by: Verified Customer WB Human 12/12/2014

Summary

• Application: Western Blot
• Sample Tested: See PMID 22277691
• Species: Human

Blocking

• Blocking Details: See PMID 22277691

Details

• Detection Notes: See PMID 22277691

Comments

• Comments: Published in PMID 22277691

Bioinformatics

• Gene Symbol: HIF1AN
• Entrez:
  • 55662(Human)
  • 29560(Rat)
• Uniprot:
  • Q9NWT6(Human)