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.
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
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
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.
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.
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