FIH-1/HIF-1AN Antibody (162c) - BSA Free

Western Blot: Factor Inhibiting HIF-1 Antibody (162c) [NBP1-30333] - Analysis in A431 cell lysates using NBP1-30333.

Immunocytochemistry/Immunofluorescence: Factor Inhibiting HIF-1 Antibody (162c) [NBP1-30333] - FIH (162C) antibody was tested in A431 cells with FITC (green). Nuclei and alpha-tubulin were counterstained with Dapi (blue) and Dylight 550 (red).

Immunohistochemistry: Factor Inhibiting HIF-1 Antibody (162c) [NBP1-30333] - Analysis of FIH in human renal cancer using DAB with hematoxylin counterstain.

• Reactivity: Hu
• Applications: WB, ICC/IF, IHC
• Clone: 162c
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

FIH-1/HIF-1AN Antibody (162c) - BSA Free Summary

• Immunogen: Full length human Factor Inhibiting HIF-1 protein [Swiss-Prot# Q9NWT6]
• Localization: Nuclear
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: HIF1AN
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:25
  • Immunohistochemistry 1:400
  • Immunohistochemistry-Paraffin 1:400
  • Western Blot 5 ug/ml
• Application Notes: This Factor Inhibiting HIF-1 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry paraffin embedded sections and Western blot analysis where a band can be seen at 40 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. 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.
• Theoretical MW: 40 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:

  Jurkat Whole Cell Lysate
  A-431 Whole Cell Lysate
  HIF-1 alpha Knockout CoCl2-treated/untreated HeLa Cell Lysate
  HIF-1 alpha Knockout Hypoxic-treated/untreated HeLa Cell Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine and 0.15M NaCl
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for FIH-1/HIF-1AN Antibody (162c) - BSA Free

• 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 (NBP1-30333)(9)

Publications using NBP1-30333

• Pickel C, Gunter J, Ruiz-Serrano A et al. Oxygen-dependent bond formation with FIH regulates the activity of the client protein OTUB1 Redox Biology 2019-07-01 [PMID: 31299612] (WB, Human)
• Ma Mingyang, Hua Shuyao, Li Gang et al. Prolyl hydroxylase domain protein 3 and asparaginyl hydroxylase factor inhibiting HIF-1 levels are predictive of tumoral behavior and prognosis in hepatocellular carcinoma. Oncotarget 2017-02-21 [PMID: 28099905] (IF/IHC, Human)
• Mysore VS, Szablowski J, Dervan PB, Frost PJ. A DNA-binding Molecule Targeting the Adaptive Hypoxic Response in Multiple Myeloma Has Potent Antitumor Activity. Mol Cancer Res. [PMID: 26801054] (WB, Human)
• Wollenick K, Hu J, Kristiansen G et al. Synthetic transactivation screening reveals ETV4 as broad coactivator of hypoxia-inducible factor signaling. Nucleic Acids Res 2012-03-01 [PMID: 22075993] (WB, Human)
• Khan MN, Bhattacharyya T, Andrikopoulos P et al. Factor inhibiting HIF (FIH-1) promotes renal cancer cell survival by protecting cells from HIF-1alpha-mediated apoptosis Br J Cancer 2011-03-29 [PMID: 21386837] (WB, Human)
• Yan M, Rayoo M, Takano EA et al. BRCA1 tumours correlate with a HIF-1alpha phenotype and have a poor prognosis through modulation of hydroxylase enzyme profile expression. Br J Cancer;101(7):1168-74. 2009-10-06 [PMID: 19724277] (IHC-P, Human)
• Cockman ME, Lancaster DE, Stolze IP, Hewitson KS, McDonough MA, Coleman ML, Coles CH, Yu X, Hay RT, Ley SC, Pugh CW, Oldham NJ, Masson N, Schofield CJ, Ratcliffe PJ. Posttranslational hydroxylation of ankyrin repeats in IkappaB proteins by the hypoxia-inducible factor (HIF) asparaginyl hydroxylase, factor inhibiting HIF (FIH). Proc Natl Acad Sci U S A;103(40):14767-72. 2006-10-03 [PMID: 17003112] (WB, Human)
• Soilleux EJ, Turley H, Tian YM et al. Use of novel monoclonal antibodies to determine the expression and distribution of the hypoxia regulatory factors PHD-1, PHD-2, PHD-3 and FIH in normal and neoplastic human tissues. Histopathology. 2005-12-01 [PMID: 16324198] (IHC-P, Human)
• Stolze IP, Tian YM, Appelhoff RJ et al. Genetic analysis of the role of the asparaginyl hydroxylase factor inhibiting hypoxia-inducible factor (FIH) in regulating hypoxia-inducible factor (HIF) transcriptional target genes [corrected]. J Biol Chem. 2004-10-01 [PMID: 15302861] (WB, ICC/IF, Human)

Review for FIH-1/HIF-1AN Antibody (NBP1-30333) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 10/23/2015

Summary

• Application: Western Blot
• Sample Tested: RPMI 8226 Cells, whole cell lysate
• Species: Human
• Lot: A1

Bioinformatics

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