FIH-1/HIF-1AN Antibody - BSA Free

Novus Biologicals | Catalog # NB100-428

Novus Biologicals
100% Guarantee

Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Knockdown Validated, Electron Microscopy

Cited:

Knockout Validated, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Electron Microscopy

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all FIH-1/HIF-1AN Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to the C-terminal region of human Factor Inhibiting HIF-1 (between residues 300 and the C-terminus). [UniProt# Q9NWT6]

Reactivity Notes

Human, rat and mouse (PMID 19720742).

Localization

Nuclear

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

40.3 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for FIH-1/HIF-1AN Antibody - BSA Free

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428]

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428]

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Analysis of astrocytes lysate from Rat brain using FIH-1/HIF-1AN antibody at 1:500 dilution with ECL detection. The antibody detected a major band at ~40 kDa position representing FIH-1/HIF-1AN protein.
Western Blot: FIH-1/HIF-1AN Antibody [NB100-428]

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428]

FIH-1-HIF-1AN-Antibody-Western-Blot-NB100-428-img0010.jpg
Immunohistochemistry: FIH-1/HIF-1AN Antibody [NB100-428]

Immunohistochemistry: FIH-1/HIF-1AN Antibody [NB100-428]

Immunohistochemistry: FIH-1/HIF-1AN Antibody [NB100-428] - Detection of FIH-1 in rat brain tissue.
Western Blot: FIH-1/HIF-1AN Antibody [NB100-428]

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428]

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Rat VSMC (vascular smoot muscle) cells treated (siFIH) or not (CTL) with a specific siRNA against FIH-1. Western blot image submitted by a verified customer review.
Knockdown Validated: FIH-1/HIF-1AN Antibody [NB100-428]

Western Blot: FIH-1/HIF-1AN Antibody [NB100-428]

FIH-1-HIF-1AN Antibody-Knockdown Validated-NB100-428-img0011.jpg
FIH-1/HIF-1AN Antibody

Immunohistochemistry: FIH-1/HIF-1AN Antibody [NB100-428] -

Immunohistochemistry: FIH-1/HIF-1AN Antibody [NB100-428] - 3D target protein expression in multicellular tumor spheroids3D expression analysis was carried out in the MCF-7, MDA-MB-231 & HBL-100 cell lines using multicellular tumor spheroids. The spheroids were cultured in spinner flasks for 1 week before fixation. Hypoxyprobe was used to examine the formation of hypoxic areas within the spheroids. NHE1, CAIX, MMP14 & FIH-1 expression was also analyzed. Incubation for an hour with PBS instead of primary antibody acted as a control. Image collected & cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.17143), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for FIH-1/HIF-1AN Antibody - BSA Free

Application
Recommended Usage

Immunohistochemistry

1:100

Immunohistochemistry-Frozen

1:50-1:200

Immunohistochemistry-Paraffin

1:50-1:200

Immunoprecipitation

1:10-1:500

Knockout Validated

reported in scientific literature (PMID 31937588)

Western Blot

1:1000-1:5000

Reviewed Applications

Read 3 reviews rated 4.3 using NB100-428 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris-Citrate/Phosphate (pH 7.0 - 8.0)

Format

BSA Free

Preservative

0.1% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: FIH-1/HIF-1AN

Factor inhibiting HIF1 (FIH1) is an asparaginyl hydroxylase enzyme that regulates HIF's transcriptional activity by hydroxylating HIF-1 alpha at 'Asp-803' in the CAD (C-terminal transactivation domain). Hydroxylation via PHDs followed by proteosome degradation is another mechanism that regulates HIF1, however, FIH1 is able to exercise the control even under severe hypoxic conditions, when PHD enzymes fail to do so. Localized mainly in cytoplasm, FIH1 functions as oxygen sensor and FIH1 interaction with NOTCH1 results in its nuclear localization whereas FIH1-ABPA3 interaction leads to its perinuclear localization. During normoxic conditions, FIH1 hydroxylates a conserved asparaginyl residue within CTAD of HIF-1 alpha/HIF-2 alpha, leading to steric clash that prevents the recruitment of the co-activators p300 and CBP. On the other hand, under hypoxic conditions, FIH1 is inactive, which results in activation of HIF-alpha signaling. FIH1 is involved in transcriptional repression through interaction with HIF1A, VHL and HDAC and hydroxylates specific Asn residues within ARD (ankyrin repeat domains) of NFKB1, NFKBIA, NOTCH1, ASB4, PPP1R12A as well as other ARD-containing proteins. FIH1 also hydroxylates Asp and His residues within ARDs of ANK1 and TNKS2, respectively. FIH1 has the ability to negatively regulate NOTCH1 activity thus accelerating myogenic differentiation, and positively regulates ASB4 activity which promotes vascular differentiation. FIH1-Mint3 binding affects FIH1's ability to modify HIF-1 alpha in an oxygen-independent manner and FIH1-Bax interaction plays inhibitory role in apoptosis.

Long Name

Factor Inhibiting HIF-1

Alternate Names

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

Entrez Gene IDs

55662 (Human); 29560 (Rat)

Gene Symbol

HIF1AN

UniProt

Q9NWT6 (Human)

Additional FIH-1/HIF-1AN Products

Product Documents for FIH-1/HIF-1AN Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for FIH-1/HIF-1AN Antibody - BSA Free

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.

Citations for FIH-1/HIF-1AN Antibody - BSA Free

Customer Reviews for FIH-1/HIF-1AN Antibody - BSA Free (3)

4.3 out of 5
3 Customer Ratings
5 Stars
33%
4 Stars
67%
3 Stars
0%
2 Stars
0%
1 Stars
0%

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • FIH-1/HIF-1AN Antibody
    Name: Darren Richard
    Application: Western Blot
    Sample Tested: Vascular Smooth Muscle Cells and Whole cell lysate
    Species: Rat
    Verified Customer | Posted 06/05/2020
    Rat VSMC cells treated (siFIH) or not (CTL) with a specific siRNA against FIH-1.
    FIH-1/HIF-1AN Antibody - BSA Free NB100-428
  • FIH-1/HIF-1AN Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cell lysates from prostate cancer cell lines
    Species: Human
    Verified Customer | Posted 11/12/2017
    Sample: Whole cell lysates of prostate cancer cell lines (20ug) Blocking: 5% nonfat dry milk reconstituted in TBS-T for 1 hour at room temperature Primary Antibody diluted (1:500) in 5% nonfat dry milk reconstituted in TBS-T overnight at 4deg C Secondary Antibody: Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution in TBS-T
    FIH-1/HIF-1AN Antibody - BSA Free NB100-428
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 22277691
    Species: Human
    Verified Customer | Posted 12/12/2014

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Protocols

View specific protocols for FIH-1/HIF-1AN Antibody - BSA Free (NB100-428):


1. Prepared 18 micron frozen rat brain tissue sections (post fixed with 10% formalin for 30 minutes).

2. Dilute the primary antibody 1:100 and incubated with diluted FIH antibody (cat# NB 100-428) at 4C, overnight.

3. Diluted the secondary antibody, biotinylated anti-rabbit IgG, according to the manufacturer's guidelines and incubated overnight at 4C.

4. Diluted the tertiary antibody, HRP-conjugated streptavidin, according to the manufacturer's guidelines and incubated overnight at 4C.

5. Visualized with DAB oxidation by HRP.


1. (30-40ug) of protein were separated via gel electrophoresis (10-20%) under denaturing conditions and transferred to nitrocellulose membrane.

2. Membrane was blocked overnight (4C) in TBS/Tween-20 (TBST) + 5% milk (blocking buffer).

3. Membrane was incubated with primary anti-FIH [cat# NB 100-428] (1:500) in blocking buffer for 2 hours at room temp.

4. Membrane was washed 3 times with TBST. 10 minutes each wash.

5. Membrane was incubated with HRP conjugated Donkey anti-rabbit secondary in blocking buffer for 2 hours at room temp.

6. Membrane was washed 3 times with TBST. 10 minutes each wash.

7. Membranes were developed for visualization using Super Signal(R) West Pico Chemiluminescent Substrate and captured on Hyperfilm ECL (Amersham Biosciences).




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FAQs

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Associated Pathways

HIF Repressor Pathways
HIF Repressor Pathway Thumbnail