Key Product Details

Species Reactivity

Human, Mouse

Applications

Western Blot, Immunocytochemistry, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 745122
View all FIH-1/HIF-1AN Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human FIH-1/HIF-1AN
Met1-Asn349
Accession # Q9NWT6

Specificity

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

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human/Mouse FIH-1/HIF-1AN Antibody

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

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 antibody in A172 Human Cell Line by Immunocytochemistry (ICC).

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.

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.
Detection of Mouse FIH-1/HIF-1AN antibody by Simple WesternTM.

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.

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

Application
Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed A172 human glioblastoma cell line and C2C12 mouse myoblast cell line

Simple Western

12.5 µg/mL
Sample: C2C12 mouse myoblast cell line

Western Blot

0.25 µg/mL
Sample: A172 human glioblastoma cell line and C2C12 mouse myoblast cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

FIH1, HIF-1AN, HIF1AN, Peptide-aspartate beta-dioxygenase

Entrez Gene IDs

55662 (Human); 319594 (Mouse); 309434 (Rat)

Gene Symbol

HIF1AN

UniProt

Q9NWT6 (Human)

Additional FIH-1/HIF-1AN Products

Product Documents for Human/Mouse FIH-1/HIF-1AN Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse FIH-1/HIF-1AN Antibody

For research use only

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Protocols

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FAQs

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Associated Pathways

HIF Repressor Pathways HIF Repressor Pathway Thumbnail