HIF-1 alpha Antibody

Images

 
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.
Western Blot: HIF-1 alpha Antibody [NB100-654] - Panc0813 human pancreatic cell line. Image supplied by customer using Lot C-2.
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
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

Summary
Reactivity Hu, Mu, Rt, PoSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml
   

Biological Strategies

     

HIF-1 alpha Antibody Summary

Immunogen
A fusion protein containing amino acids 432 - 528 of human HIF-1 alpha [UniProt# Q16665].
Localization
Nuclear
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
HIF1A
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • 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. Separated by Size-Wes, Sally Sue/Peggy Sue.
Theoretical MW
93 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.
Positive Control
COS-7 Lysate (NB800-PC26)
Placenta Lysate (NB820-59248)
Bone Marrow Lysate (NB820-59283)
Reviewed Applications
Read 1 Review rated 5
using
NB100-654 in the following applications:

Publications
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).

Packaging, Storage & Formulations

Storage
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Buffer
Tris-Glycine and 0.15M NaCl
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

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
  • HIF 1A
  • HIF1 alpha
  • HIF-1 alpha
  • HIF1A
  • HIF-1a
  • 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
  • MOP1HIF1-ALPHA
  • PAS domain-containing protein 8
  • PASD8alpha subunit (basic helix-loop-helix transcriptionfactor)

Background

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.

Limitations

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)(9)

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

We have publications tested in 5 applications: ICC/IF, IHC, IHC-Fr, IHC-P, WB.


Filter By Application
ICC/IF
(1)
IHC
(1)
IHC-Fr
(1)
IHC-P
(2)
WB
(2)
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Human
(4)
Mouse
(3)
Rat
(1)
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Showing Publications 1 - 9 of 9.
Publications using NB100-654 Applications Species
Bouchard G, Therriault H, Geha S et al. Radiation-induced lung metastasis development is MT1-MMP-dependent in a triple-negative breast cancer mouse model. Br J Cancer 2017 Feb 14 [PMID: 28103615] (Mouse) Mouse
Knudsen ES, Balaji U, Freinkman E et al. Unique metabolic features of pancreatic cancer stroma: relevance to the tumor compartment, prognosis, and invasive potential. Oncotarget. Nov 29 2016 [PMID: 27623078] (IHC, Human) IHC Human
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)

Details:
HIF1 alpha antibody used for WB on cytoplasmic and nuclear extractions of normoxia, hypoxia or uM 150 CoCl2 exposed primary satellite cells which were isolated from male Sprague Dawley rats (Figure 4).
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]

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

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

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

ApplicationWestern Blot
Sample Testedhuman cancer lines
SpeciesHuman
LotC-2
CommentsThis antibody is very easy to works with. It gives strong and clean signals in my experiment.

Blocking

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

Details

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

Comments

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 405 Labeled NB100-654AF405
Alexa Fluor 488 Labeled NB100-654AF488
Alexa Fluor 647 Labeled NB100-654AF647
Alexa Fluor 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 27 blog posts - Show all blog posts.

Forecasting and Targeting a Rare Cancer with Hypoxia-Inducible Factor
By Jamshed Arslan Pharm.D. Cancers of nerve, adipose, and other soft tissues are called soft tissue sarcomas (STS). Malignant peripheral nerve sheath tumor (MPNST) is an example of a rare and hard-to-treat STS; eve...  Read full blog post.

The role of HIF-1 Alpha signaling in the retina under hypoxic conditions
Hypoxia inducible factor 1 (HIF-1) is a protein that plays an essential role in hypoxia, or low levels of cellular oxygen. HIF-1 is a heterodimeric protein that consists of a constitutively expressed beta subunit and oxygen related alpha subunit. ...  Read full blog post.

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.

Showing 1-10 of 27 blog posts - Show all blog posts.

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Recent Reviews

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Ming Lu
07/25/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol HIF1A
Entrez