HIF-1 alpha Antibody


Western Blot: HIF-1 alpha Antibody [NB100-654] - Detection of HIF-1 alpha in Cobalt Chloride treated/untreated Cos-7 nuclear extracts using NB100-654.
Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-654] - IHC analysis of a formalin-fixed paraffin-embedded tissue section of human endometrium carcinoma AN3CA cell line based xenograft using rabbit polyclonal ...read more
Western Blot: HIF-1 alpha Antibody [NB100-654] - Panc0813 human pancreatic cell line. Image supplied by customer using Lot C-2.
Simple Western: HIF-1 alpha Antibody [NB100-654] - Simple Western lane view shows a specific band for HIF-1 alpha in 0.5 mg/ml of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the ...read more

Product Details

Reactivity Hu, Mu, Rt, PoSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC, IHC-Fr, IHC-P
1.0 mg/ml

HIF-1 alpha Antibody Summary

A fusion protein containing amino acids 432 - 528 of human HIF-1 alpha [UniProt# Q16665].
Immunogen affinity purified
Innovator's Reward
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Packaging, Storage & Formulations

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Tris-glycine, 150 mM NaCl
0.05% Sodium Azide
1.0 mg/ml
Immunogen affinity purified


  • Western Blot 1:1000
  • Simple Western 1:100
  • Immunocytochemistry/Immunofluorescence 1:10 - 1:500
  • Immunohistochemistry 1:100 - 1:300
  • Immunohistochemistry-Frozen 1:100 - 1:300
  • Immunohistochemistry-Paraffin 1:100 - 1:300
Application Notes
Nuclear extracts are recommended. ICC/IF, IHC-P and IHC-F usages were reported in scientific literature. In Simple Western only 10 - 15 ul of the recommended dilution is used per data point.
Theoretical MW
93 kDa
Reviewed Applications
Read 1 Review rated 5
NB100-654 in the following applications:

Read Publications using
NB100-654 in the following applications:

Reactivity Notes

Immunogen sequence has 93% homology to Bovine and 90% homology to Mouse. Mouse reactivity was reported in scientific literature (PMID: 23959856). Rat reactivity was reported in scientific literature (PMID: 24122166).

Alternate Names for HIF-1 alpha Antibody

  • ARNT-interacting protein
  • Basic-helix-loop-helix-PAS protein MOP1
  • BHLHE78
  • Class E basic helix-loop-helix protein 78
  • HIF1 alpha
  • HIF-1 alpha
  • HIF1A
  • HIF-1-alpha
  • HIF1-alpha
  • hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcriptionfactor)
  • hypoxia-inducible factor 1-alpha
  • Member of PAS protein 1
  • member of PAS superfamily 1
  • PAS domain-containing protein 8
  • PASD8alpha subunit (basic helix-loop-helix transcriptionfactor)


HIF1 (hypoxia-inducible factor 1), a heterodimeric transcription factor complex central to cellular response to hypoxia, consists of two subunits (HIF-1 alpha and HIF-1 beta) which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family. Expression of HIF-1 alpha protein is regulated by cellular oxygen level alterations as well as in oxygen-independent manner via different cytokines (through the PI3K-AKT-mTOR pathway), growth factors, oncogenic activation, or loss of tumor suppressor function etc. In normoxic cells, HIF-1 alpha is proline hydroxylated leading to a conformational change that promotes its binding to the VHL (von Hippel Lindau) protein E3 ligase complex; ubiquitination and followed by rapid proteasomal degradation. Hypoxia as well as chemical hydroxylase inhibitors (desferrioxamine, cobalt etc.) inhibit HIF-1 alpha degradation and lead to its accumulation in the cells, whereas, contrastingly, HIF-1 beta/ARNT (AhR nuclear translocator) remains stable under both conditions. Besides their critical role in hypoxic response, HIF1s regulates the transcription of genes responsible for angiogenesis, erythropoiesis/iron-metabolism, glucose metabolism, cell proliferation/survival, adipogenesis, carotid body formation, B lymphocyte development and immune reactions.


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.

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Publications for HIF-1 alpha Antibody (NB100-654)(7)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 4 applications: ICC/IF, IHC-Fr, IHC-P, WB.

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Showing Publications 1 - 7 of 7.
Publications using NB100-654 Applications Species
De Bruycker S, Vangestel C, Van den Wyngaert T et al. Baseline [(18)F]FMISO microPET as a Predictive Biomarker for Response to HIF-1alpha Inhibition Combined with 5-FU Chemotherapy in a Human Colorectal Cancer Xenograft Model. Mol Imaging Biol. 2016 Jan 04 [PMID:26728163] (IHC-P, Human) IHC-P Human
Flann KL, Rathbone CR, Cole LC et al. Hypoxia simultaneously alters satellite cell-mediated angiogenesis and hepatocyte growth factor expression. J Cell. Physiol. 2014 May 01 [PMID:24122166] (WB, Rat) WB Rat
Cao Y, Eble JM, Moon E et al. Tumor Cells Upregulate Normoxic HIF-1alpha in Response to Doxorubicin. Cancer Res. 2013 Aug 19 [PMID:23959856] (WB, Human, Mouse) WB Human, Mouse
Chiang C-K, Tanaka T, Inagi R et al. Indoxyl sulfate, a representative uremic toxin, suppresses erythropoietin production in a HIF-dependent manner. Lab Invest. 2011 Aug 22. [PMID:21863063]
Thiersch M, Lange C, Joly S et al. Retinal neuroprotection by hypoxic preconditioning is independent of hypoxia-inducible factor-1 alpha expression in photoreceptors. Eur J Neurosci 2009 Jun [PMID:19508692] (IHC-Fr, ICC/IF, Mouse) IHC-Fr, ICC/IF Mouse
Yasui H, Ogura A, Asanuma T et al. Inhibition of HIF-1alpha by the anticancer drug TAS106 enhances X-ray-induced apoptosis in vitro and in vivo. Br J Cancer;99(9):1442-52. 2008 Nov 4. [PMID:18854835] (IHC-P, Human) IHC-P Human
Mojsilovic-Petrovic J, Callaghan D, Cui H et al. Hypoxia-inducible factor-1 (HIF-1) is involved in the regulation of hypoxia-stimulated expression of monocyte chemoattractant protein-1 (MCP-1/CCL2) and MCP-5 (Ccl12) in astrocytes. J Neuroinflammation 2007 [PMID:17474992]

Reviews for HIF-1 alpha Antibody (NB100-654) (1) 51

Average Rating: 5
(Based on 1 reviews)
We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: WB.

Reviews using NB100-654:
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Images Ratings Applications Species Date Details
Western Blot HIF-1 alpha NB100-654
reviewed by:
WB Human 07/25/2014


ApplicationWestern Blot
Sample Testedhuman cancer lines
CommentsThis antibody is very easy to works with. It gives strong and clean signals in my experiment.


Blocking Details2% cold fish gelantin

Primary Anitbody

Dilution Ratio1:1000, overnight at 4C in blocking buffer

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG, HRP
Secondary Manufacturer Cat#Cell Signaling Anti-rabbit IgG, HRP-linked Antibody #7074
Secondary Concentration1:5000


Detection Notes30 second exposure will give strong signal, negative controls are cells grew in normal condition (5% CO2)


CommentsThis antibody is very easy to works with. It gives strong and clean signals in my experiment.

Product General Protocols

View specific protocols for HIF-1 alpha Antibody (NB100-654): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for HIF-1 alpha Antibody (NB100-654). (Showing 1 - 4 of 4 FAQs).

  1. Why is there a difference between the theoretical MW for HIF1A and the observed MW for HIF-1 alpha?
    • HIF1A, like many other proteins, has post-translational modifications. Depending on the size, amount and nature of the post-translational modifications, it can cause subtle to very large changes in molecular weight.
  2. Which antibody(ies) do you recommend for the detection of HIF-1a by immunohistochemistry in the sections of paraffin-embedded mouse liver samples? I would appreciate if you can give me several choices and rank them in the order of performance. My goal is to distinguish HIF upregulation by prolyl hydroxylase inhibitor in different liver cells.
    • All of our antibodies are of high quality and are well tested/validated in species/applications we list on the datasheet. However, as desired by you, I am suggesting you four HIF-1 alpha antibodies based upon customer reviews, as well as the number of peer reviewed publications in which these products has been cited by researchers from reputed institutes. (1) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-105) (cited in at least 218 peer reviewed publications) (2) HIF-1 alpha Antibody (cat# NB100-479) (cited in at least 51 peer reviewed publications) (3) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-123 ) (cited in at least 38 peer reviewed publications) (4) HIF-1 alpha Antibody (cat# NB100-449 ) (cited in at least 31 peer reviewed publications).
  3. I would like to know, does it exist path for detection HIF 1 in the venous blood before and after revascularization of the leg?
    • We are not entirely sure if HIF-1 alpha will be present in the leg after revascularization. It may be present but you may want to search the literature to see if this has been looked at before. If not, then this would certainly be an experiment worth doing.
  4. What is the molecular weight (kDa) of protein HIF 1 alpha in western blot?
    • The theoretical molecular weight of HIF 1-alpha is ~93kDa, however, you will likely see a band between 100-120kDa due to phosphorylation.

Positive Control Lysate(s)

Secondary Antibodies


Isotype Controls

Other Available Formats

Alexa Fluor (R) 405 Labeled NB100-654AF405
Alexa Fluor (R) 488 Labeled NB100-654AF488
Alexa Fluor (R) 647 Labeled NB100-654AF647
Alexa Fluor (R) 700 Labeled NB100-654AF700
Biotin Labeled NB100-654B
DyLight 405 Labeled NB100-654V
DyLight 488 Labeled NB100-654G
DyLight 550 Labeled NB100-654R
DyLight 650 Labeled NB100-654C
FITC Labeled NB100-654F
HRP Labeled NB100-654H

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Blogs on HIF-1 alpha. Showing 1-10 of 25 blog posts - Show all blog posts.

The relationship between Ki67 and HIF-1 in cancer
Ki67, also known as MKI67, is best known as the leading marker of cellular proliferation. Ki67 is regulated by a balance between synthesis and degradation, and often carries a very short half-life.  First discovered to be located to dividing cells,...  Read full blog post.

Controls for HIF-1 Alpha's WB, ICC-IF, IHC, IP & FLOW Analysis
Tips on positive and negative controls for HIF-1 alpha antibodies is one of the most Frequently Asked Questions on Hypoxia and HIFs. Here are top 5 suggestions from Novus Biologicals: The degradation of HIF1 alpha is the most common issue whic...  Read full blog post.

Understanding the relationship between HIF-1 alpha, Hypoxia and Epithelial-Mesenchymal Transition
Epithelial-mesenchymal transition (EMT) is a natural process by which epithelial cells lose their polarity and intercellular adhesion, and gain the migratory invasive properties of mesenchymal stem cells that can differentiate into a variety of cel...  Read full blog post.

HIF-2 alpha: HIF1A's Homologue with Similar and Divergent Functions
HIF-2 alpha is a member of the heterodimeric hypoxia-inducible factors/HIFs family (HIF-1, HIF-2, and HIF-3) which contains a common beta subunit but differ in their alpha subunits. Also called as EPAS1 or Mop2, HIF-2 alpha regulates cellular adapt...  Read full blog post.

HIF-3 alpha: a versatile target with hypoxia dependent and independent functions
By: Subhash GangarHIF-3 alpha (hypoxia-inducible factor 3-alpha/ HIF3A) represents an isoform of HIF-alpha subunits which heterodimerize with stable beta subunit (HIF-beta) for the regulation of HIF target genes through binding to hypoxia respon...  Read full blog post.

HIF-1 alpha - sensing and responding to changing oxygen levels
Hypoxia-inducible factor 1 (HIF-1) allows cells to respond to changing levels of oxygen in the environment. HIF-1 is a heterodimeric transcription factor consisting of alpha and beta subunits. Under normal conditions HIF-1 alpha is continuously sy...  Read full blog post.

Comprehensive Autophagy Research Tools - New Catalog Available Now!
Autophagy, a protein degradation process through autophagosome-lysosomal pathway, is important for cellular homeostasis and plays a role in many diseases. To help researchers learn more about this process and the products available for its study, N...  Read full blog post.

HIF-1 beta: A dimerization partner of HIF-1 alpha required for an adaptive response to hypoxia
Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. Hypoxia-indu...  Read full blog post.

HIF-1 Alpha: Infographic
Encoded by the HIF1A gene, HIF-1 alpha has a critical role in cellular response to hypoxia. In hypoxic conditions, HIF-1 alpha activates the transcription of several genes to facilitate metabolic reaction for lack of oxygen. In normoxic conditions, HI...  Read full blog post.

Controlling the HIF-1 Switch
Hypoxia-inducible factor-1 is a major transcription factor composed of two subunits: HIF-1alpha and HIF-1 beta. Under normoxic conditions, HIF-1 alpha is targeted to proteosomal degradation via ubiquitination. On the other hand during hypoxic conditio...  Read full blog post.

Showing 1-10 of 25 blog posts - Show all blog posts.

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Gene Symbol HIF1A

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