FIH-1/HIF-1AN Antibody (745122) [Unconjugated]

Western blot shows lysates of A172 human glioblastoma cell line and C2C12 mouse myoblast cell line. PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human/Mouse FIH-1/ HIF-1AN Monoclonal Antibody (Catalog # MAB7874) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for FIH-1/HIF-1AN at approximately 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

FIH-1/HIF-1AN was detected in immersion fixed A172 human glioblastoma cell line using Mouse Anti-Human/Mouse FIH-1/HIF-1AN Monoclonal Antibody (Catalog # MAB7874) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western lane view shows lysates of C2C12 mouse myoblast cell line, loaded at 0.5 mg/mL. A specific band was detected for FIH-1/HIF-1AN at approximately 40 kDa (as indicated) using 12.5 µg/mL of Mouse Anti-Human/Mouse FIH-1/HIF-1AN Monoclonal Antibody (Catalog # MAB7874). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

FIH-1/HIF-1AN was detected in immersion fixed C2C12 mouse myoblast cell line using Mouse Anti-Human/Mouse FIH-1/HIF-1AN Monoclonal Antibody (Catalog # MAB7874) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, ICC/IF
• Clone: 745122
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

FIH-1/HIF-1AN Antibody (745122) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human FIH-1/HIF-1AN
  Met1-Asn349
  Accession # Q9NWT6
• Specificity: Detects human FIH-1/HIF-1AN in ELISAs.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: HIF1AN
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-25 ug/mL
  • Simple Western 12.5 ug/mL
  • Western Blot 0.25 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for FIH-1/HIF-1AN Antibody (745122) [Unconjugated]

• 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 hypoxia-inducible factor 1 (FIH-1), also known as HIF-1AN, is a 40-45 kDa oxygenase that hydroxylates asparagine, aspartate, and histidine residues in target proteins. Its homodimerization is required for substrate recognition. Under normoxic conditions, FIH-mediated hydroxylation inhibits the function of HIF-1 alpha and HIF-2 alpha by preventing their association with the transcriptional coactivator p300. FIH activity is inhibited under hypoxic conditions which allows HIF to mediate hypoxia-induced gene transcription. FIH also hydroxylates amino acids within the ankyrin repeat domains (ARD) of various other proteins including IkB-alpha, NFkB1, AnkyrinR, AnkyrinB, Tankyrase 2, MYPT1 smooth muscle myosin phosphatase, and Notch. Human FIH-1 shares 97% aa sequence identity with mouse and rat FIH-1.

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.

Bioinformatics

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