HIF-1 alpha Antibody (ESEE122)

Images

 
Simple Western: HIF-1 alpha Antibody (ESEE122) [NB100-131] - 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 ...read more
Western Blot: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Western Blot of HIF1a in Cos7 cells treated or untreated with CoCl2.
Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Detection of HIF-1 alpha (red dye 568) in a cultured raw mouse macrophage cell line, using NB100-131. Photos courtesy of Susan ...read more
Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (ESEE122) [NB100-131] - HIF-1 Alpha staining in hypoxia-induced human placenta.
Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - IHC analysis of a formalin fixed paraffin embedded tissue section of human renal cancer xenograft using Lot 83115 of HIF1 alpha antibody at ...read more
Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Detection of HIF-1 alpha (red dye) in a cell cytospin from a lavage of a murine skin pouch infected with Staph Aureus, using ...read more
Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Detection of HIF-1 alpha (red dye 568) in a cultured raw mouse macrophage cell line, using NB100-131. 100X magnification. Photos ...read more
Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Detection of HIF-1 alpha (red dye) in a cell cytospin from a lavage of a murine skin pouch infected with Staph Aureus, using ...read more
Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - IHC analysis of a formalin fixed paraffin-embedded mouse kidney tissue section using HIF-1 alpha antibody clone ESEE122 at 1ug/ml ...read more
Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - analysis of HIF-1 alpha in paraffin-embedded mouse kidney tissue section using anti-HIF-1 alpha antibody. Image from verified customer review.
Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Negative control stain of human placenta (from sea level) using mouse IgG at 1:100. 4uM paraffin-embedded section.

Product Details

Summary
Reactivity Hu, Mu, Rt, Bv, CaSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC, IHC-Fr, IHC-P, IP
Clone
ESEE122
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1.0 mg/ml
   

Biological Strategies

     

HIF-1 alpha Antibody (ESEE122) Summary

Immunogen
Human HIF-1 alpha, corresponding to amino acids 329 - 530 [UniProt# Q16665].
Localization
Cytoplasm, Nucleus
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
HIF1A
Purity
Protein G 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 1:500 - 1:1000
  • Simple Western 1:2000
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:100 - 1:5000
  • Immunohistochemistry-Frozen 1:100 - 1:5000
  • Immunohistochemistry-Paraffin 1:100 - 1:5000
  • Immunoprecipitation 1:10 - 1:500
Application Notes
Variable results have been obtained in Western blot.

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
Kidney Lysate (NB820-59231)
COS-7 Lysate (NB800-PC26)
Placenta Lysate (NB820-59248)
Reviewed Applications
Read 3 Reviews rated 4.3
using
NB100-131 in the following applications:

Publications
Read Publications using
NB100-131 in the following applications:

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

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
Protein G purified

Alternate Names for HIF-1 alpha Antibody (ESEE122)

  • 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 (NB100-131)(63)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Bovine, Canine.

We have publications tested in 7 applications: FLOW, ICC/IF, IHC, IHC-Fr, IHC-P, IHC-P,IHC-Fr, WB.


Filter By Application
FLOW
(2)
ICC/IF
(7)
IHC
(17)
IHC-Fr
(1)
IHC-P
(19)
IHC-P,IHC-Fr
(1)
WB
(10)
All Applications
Filter By Species
Human
(28)
Mouse
(7)
Rat
(5)
Bovine
(2)
Canine
(2)
All Species
Showing Publications 1 - 10 of 63. Show All 63 Publications.
Publications using NB100-131 Applications Species
Gustafsson H, Kale A, Dasu A et al. EPR Oximetry of Cetuximab-Treated Head-and-Neck Tumours in a Mouse Model. Cell Biochem. Biophys. 2017 Jul 29 [PMID: 28756482] (Mouse) Mouse
Boyer DF, McKelvie PA, de Leval L et al. Fibrin-associated EBV-positive Large B-Cell Lymphoma: An Indolent Neoplasm With Features Distinct From Diffuse Large B-Cell Lymphoma Associated With Chronic Inflammation. Am. J. Surg. Pathol. 2017 Mar 01 [PMID: 28195879] (IHC) IHC
Huang T, Chung S, Chua S et al. Effect of early administration of lower dose versus high dose of fresh mitochondria on reducing monocrotaline-induced pulmonary artery hypertension in rat. Am J Transl Res. Dec 30 2016 [PMID: 28077992] (ICC/IF, Rat) ICC/IF Rat
Hjelmgren O, Gellerman K, Kjelldahl J et al. Increased Vascularization in the Vulnerable Upstream Regions of Both Early and Advanced Human Carotid Atherosclerosis. PLoS ONE. Dec 15 2016 [PMID: 27973542] (IHC, Human) IHC Human
Cahill LS, Gazdzinski LM, Tsui AK et al. Functional and anatomical evidence of cerebral tissue hypoxia in young sickle cell anemia mice. J. Cereb. Blood Flow Metab. May 10 2016 [PMID: 27165012] (IHC-P, Mouse) IHC-P Mouse
Koh J, Jang JY, Keam B et al. EML4-ALK enhances programmed cell death-ligand 1 expression in pulmonary adenocarcinoma via hypoxia-inducible factor (HIF)-1alpha and STAT3. Oncoimmunology 2016 Mar [PMID: 27141364] (IHC) IHC
Miller AN, Deal D, Green J et al. Use of the Reamer/Irrigator/Aspirator Decreases Carotid and Cranial Embolic Events in a Canine Model. J Bone Joint Surg Am. 2016 Apr 20 [PMID: 27098324] (IHC, Canine) IHC Canine
Fan Y, Li H, Ma X et al. Dicer suppresses the malignant phenotype in VHL-deficient clear cell renal cell carcinoma by inhibiting HIF-2alpha. Oncotarget Apr 5 2016 [PMID: 26943772] (WB, Human) WB Human
Kim Y, Nam HJ, Lee J et al. Methylation-dependent regulation of HIF-1alpha stability restricts retinal and tumour angiogenesis. Nat Commun. 2016 Jan 13 [PMID: 26757928]
Ochi F, Shiozaki A, Ichikawa D et al. Carbonic Anhydrase XII as an Independent Prognostic Factor in Advanced Esophageal Squamous Cell Carcinoma. J Cancer. 2015 Aug 28 [PMID: 26316888] (IHC-P, Human)

Details:
HIF-1 alpha antibody (clone ESEE122) was used for IHC-P analysis of tissue sections from human subjects with advanced esophageal squamous cell carcinoma (Fig 2).
IHC-P Human
Show All 63 Publications.

Reviews for HIF-1 alpha Antibody (NB100-131) (3) 4.33

Average Rating: 4.3
(Based on 3 reviews)
We have 3 reviews tested in 2 species: Human, Mouse.
We have 3 reviews tested in 1 application: IHC-P.

Reviews using NB100-131:
Filter by Applications
IHC-P
(2)
All Applications
Filter by Species
Human
(1)
Mouse
(2)
All Species
Images Ratings Applications Species Date Details
 HIF-1 alpha NB100-131
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reviewed by:
Anonymous
Mouse 02/04/2015
View

Summary

SpeciesMouse
FileView PDF
Immunohistochemistry-Paraffin HIF-1 alpha NB100-131
Enlarge
reviewed by:
Anonymous
IHC-P Mouse 01/31/2013
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedMurine Kidney
SpeciesMouse
LotG5
CommentsExcellent antibody providing strong positive staining with low levels of background, highly recommended for IHC-P, thanks Novus!

Blocking

Blocking DetailsBlocked with serum free protein block

Primary Anitbody

Dilution RatioIncubated for 1 hour RT at a concentration of 1ug/ml

Secondary Antibody

Secondary DescriptionX-cell plus universal HRP polymer detection system (Menarini Diagnostics)
Secondary Manufacturer Cat#MP-XCP-U25
Secondary ConcentrationAccording to manufacturers instructions

Details

Detection NotesVector SG chromagen substrate (5 minute incubation)
Fixation DetailsTissue fixed in formyl saline, heat mediated antigen retrieval performed at 100C in citrate (pH6) buffer
Wash Description4 x 5 minute PBS washes

Comments

CommentsExcellent antibody providing strong positive staining with low levels of background, highly recommended for IHC-P, thanks Novus!
 
reviewed by:
Anonymous
IHC-P Human 02/02/2009
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample Testedhuman melanoma
SpeciesHuman
LotE
CommentsAt this dilution there is still some background but the nuclear staining is very clear.

Blocking

Blocking DetailsBlocking Buffer: goat serum, Blocking Time: 30 minutes, Blocking Temp: room temperature

Primary Anitbody

Dilution RatioDilution Ratio: 1/1000, Incubation Dilution Buffer: blocking buffer, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

Secondary DescriptionSecondary Ab: goat anti rabbit from Santa Cruz, Secondary Ab Dilution: 1/200

Comments

CommentsAt this dilution there is still some background but the nuclear staining is very clear.

Product General Protocols

View specific protocols for HIF-1 alpha Antibody (NB100-131): 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 (NB100-131). (Showing 1 - 8 of 8 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.
  5. Our customer wants to buy the peptide for your antibody NB100-131. Which catalog number does the peptide have?
    • This antibody was made to a recombinant protein containing amino acids 329-530 of HIF-1 alpha. Unfortunately we do not carry the recombinant protein as a sellable item.
  6. Do you have any more protocol information regarding the cytospin staining images on your datasheet, credited to Susan Alexander and Hattie Gresham?
    • I do have the following notes: 1.) On cultured raw cells (mouse macrophage cell line). 2.) On a cell cytospin from a lavage of a skin pouch (mouse) infected with staph aureus. 10 100X pic 3.) Also on a cell cytospin from a lavage of a skin pouch of a mouse infected with staph aureus. 14. WT pic. 4.) on cultured raw cells. 19. pic The blue staining is DAPI (nuclear stain). I conjugated the stain with a kit from molecular probes (A-20184) & simply followed their protocol. I am uncertain of the final antibody concentration. It may possibly be 25 ug/mL, and I am using it at 1:2 or 1:3 (about 8-12ug/mL). I don't have a protocol written out yet.
  7. Is your HIF-1 alpha Antibody able to work in Western Blot application?
    • Yes, this antibody can be used in Western Blot.
  8. I've done colorations with an HIF-1alpha antibody and am now wondering if it is HIF-1alpha we see or it may be cross-reactivity with any other in the cytoplasm in mast cells. The image is reminiscent of the Immunofluorescence staining available on your site for HIF-1 alpha Antibody (ESEE122) [NB100-131] (Detection of HIF-1 alpha (red dye) in a cell cytospin from a lavage of a murine skin pouch infected with Staph Aureus, using NB100-131.) Gresham and Alexander have done that picture, but I cannot find it published. How to explain the appearance of HIF-1 staining. Our staining is localized to the nucleus, but is bead-like.
    • HIF-1 alpha can be found at very low levels in the cytoplasm under normoxic conditions. During hypoxia, HIF1a is stabilized and translocates to the nucleus to act as a transcription factor. You should expect to see nuclear staining in your hypoxic samples, but may also observe faint cytoplasmic staining corresponding to degraded HIF. We do not have a publication from Gresham and Alexander on our website, and I believe that their images were submitted as a customer review of the antibody. However we do have five other publications referring to immunofluorescent staining, which you may find helpful. You can see these at the bottom of the "Images, Reviews and Publications" tab by filtering on application and selecting "ICC/IF".

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB100-131AF405
Alexa Fluor 488 Labeled NB100-131AF488
Alexa Fluor 647 Labeled NB100-131AF647
Alexa Fluor 700 Labeled NB100-131AF700
Biotin Labeled NB100-131B
DyLight 350 Labeled NB100-131UV
DyLight 405 Labeled NB100-131V
DyLight 488 Labeled NB100-131G
DyLight 550 Labeled NB100-131R
DyLight 650 Labeled NB100-131C
DyLight 680 Labeled NB100-131FR
DyLight 755 Labeled NB100-131IR
FITC Labeled NB100-131F
HRP Labeled NB100-131H

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Blogs on HIF-1 alpha. Showing 1-10 of 26 blog posts - Show all blog posts.

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.

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.

Showing 1-10 of 26 blog posts - Show all blog posts.

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Recent Reviews

4.3
5
1
4
2
3
0
2
0
1
0

Anonymous
02/04/2015
Application:
Species: Mouse

Anonymous
01/31/2013
Application: IHC-P
Species: Mouse

 
Anonymous
02/02/2009
Application: IHC-P
Species: Human

Bioinformatics

Gene Symbol HIF1A
Entrez