Human/Mouse FIH-1/HIF-1AN Antibody

    
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  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human FIH-1/HIF-1AN in ELISAs.
  • Source
    Monoclonal Mouse IgG2B Clone # 745122
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human FIH-1/HIF-1AN
    Met1-Asn349
    Accession # Q9NWT6
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.25 µg/mL
    See below
  • Simple Western
    12.5 µg/mL
    See below
  • Immunocytochemistry
    3-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human and Mouse FIH-1/HIF-1AN by Western Blot. 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.
Immunocytochemistry
FIH-1/HIF-1AN in A172 Human Cell Line. 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.
Detection of Mouse FIH-1/HIF-1AN by Simple WesternTM.

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.

Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
    Reconstitution Buffer Available
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: FIH-1/HIF-1AN
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.
  • Long Name:
    Factor Inhibiting HIF-1
  • Entrez Gene IDs:
    55662 (Human); 319594 (Mouse); 309434 (Rat)
  • Alternate Names:
    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
Related Research Areas

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