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

Catalog #: MAB7874 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB7874
MAB7874-SP

Detection of Human and Mouse FIH-1/HIF-1AN by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

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

Species Reactivity

Human, Mouse

Specificity

Detects human FIH-1/HIF-1AN in ELISAs.

Source

Monoclonal Mouse IgG_2B 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 either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

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.

Scientific Data

Western Blot

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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

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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.

Immunocytochemistry

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FIH-1/HIF-1AN in C2C12 Mouse Cell Line. 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.

Simple Western

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Detection of Mouse FIH-1/HIF-1AN by Simple Western^TM. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

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

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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