HIF-1 alpha Antibody [Unconjugated]

Images

 
Western blot shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 150 μM CoCl2 for 16 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat HIF‑1 alpha ...read more
Mouse primary kidney cells treated with 150 μM CoCl2 for overnight were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HIF‑1 alpha /DNA complexes were immunoprecipitated ...read more
Simple Western lane view shows lysates of A549 human lung carcinoma cell line untreated (-) or treated (+) with Hypoxia (1% O2), loaded at 0.2 mg/mL. A specific band was detected for HIF‑1 alpha /HIF1A at ...read more
HIF‑1 alpha /HIF1A was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with DFO but is not detected in HIF‑1 alpha /HIF1A knockout (KO) HeLa Human Cell Line cell line ...read more

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, Simple Western, IP, ChIP, ICC/IF, KO
Clonality
Polyclonal
Host
Goat
Conjugate
Unconjugated
Concentration
LYOPH

Order Details

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Catalog# & Formulation Size Price

HIF-1 alpha Antibody [Unconjugated] Summary

Immunogen
E. coli-derived recombinant human HIF-1 alpha /HIF1A
Arg575-Asn826
Accession # Q16665.1
Specificity
Detects human, mouse and rat HIF-1 alpha /HIF1A.
Source
N/A
Isotype
IgG
Clonality
Polyclonal
Host
Goat
Gene
HIF1A
Purity Statement
Antigen 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 0.5 ug/mL
  • Simple Western 5 ug/mL
  • Immunoprecipitation 2 ug/500 ug cell lysate
  • Chromatin Immunoprecipitation (ChIP) 5 ug/5 x 10^6 cells
  • Immunocytochemistry 0.3-15 ug/mL
  • Knockout Validated 0.3-15 ug/mL
Application Notes
In Simple Western only 10-15 uL of the recommended dilution is used per data point.
Reviewed Applications
Read 2 Reviews rated 4
using
AF1935 in the following applications:

Publications
Read Publications using
AF1935 in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Preservative
No Preservative
Concentration
LYOPH
Reconstitution Instructions
Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for HIF-1 alpha Antibody [Unconjugated]

  • AINT
  • anti-HIF-1 alpha
  • anti-HIF1A
  • ARNT interacting protein
  • 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
  • HIF1
  • HIF1A
  • HIF-1a
  • HIF-1alpha
  • HIF-1-alpha
  • HIF1-alpha
  • hypoxia inducible factor 1 alpha subunit, hypoxia inducible factor 1 subunit alpha
  • hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
  • hypoxia-inducible factor 1-alpha
  • Member of PAS protein 1
  • member of PAS superfamily 1
  • MOP1
  • PAS domain-containing protein 8
  • PASD8

Background

The hypoxia-inducible transcription factor 1 alpha (HIF-1 alpha ) is the regulated member of the transcription factor heterodimer HIF-1. HIF-1 binds to hypoxia-response elements (HREs) in the promoters of many genes involved in adapting to an environment of insufficient oxygen or hypoxia. Hypoxic tissue environments occur in vascular and pulmonary diseases as well as cancer, which illustrates the broad impact of gene regulation by HIF-1 alpha.

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 (AF1935)(21)

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

We have publications tested in 4 applications: ChIP, IHC, IHC-F, Western Blot.


Filter By Application
ChIP
(4)
IHC
(1)
IHC-F
(1)
Western Blot
(15)
All Applications
Filter By Species
Human
(12)
Mouse
(8)
Rat
(1)
All Species
Showing Publications 1 - 10 of 21. Show All 21 Publications.
Publications using AF1935 Applications Species
M Felsenstei, A Blank, AD Bungert, A Mueller, A Ghori, I Kremenetsk, O Rung, T Broggini, K Turkowski, L Scherschin, J Raggatz, P Vajkoczy, S Brandenbur CCR2 of Tumor Microenvironmental Cells Is a Relevant Modulator of Glioma Biology Cancers (Basel), 2020;12(7):. 2020 [PMID: 32668709] (IHC-F, Mouse) IHC-F Mouse
K Tawara, H Scott, J Emathinger, A Ide, R Fox, D Greiner, D LaJoie, D Hedeen, M Nandakumar, AJ Oler, R Holzer, C Jorcyk Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion Transl Oncol, 2018;12(2):245-255. 2018 [PMID: 30439625] (Western Blot, Human) Western Blot Human
AN Billin, SE Honeycutt, AV McDougal, JP Kerr, Z Chen, JM Freudenber, DK Rajpal, G Luo, HF Kramer, RS Geske, F Fang, B Yao, RV Clark, J Lepore, A Cobitz, R Miller, K Nosaka, AC Hinken, AJ Russell HIF prolyl hydroxylase inhibition protects skeletal muscle from eccentric contraction-induced injury Skelet Muscle, 2018;8(1):35. 2018 [PMID: 30424786] (Western Blot, Mouse) Western Blot Mouse
Q Yang, J Xu, Q Ma, Z Liu, V Sudhahar, Y Cao, L Wang, X Zeng, Y Zhou, M Zhang, Y Xu, Y Wang, NL Weintraub, C Zhang, T Fukai, C Wu, L Huang, Z Han, T Wang, DJ Fulton, M Hong, Y Huo PRKAA1/AMPK?1-driven glycolysis in endothelial cells exposed to disturbed flow protects against atherosclerosis Nat Commun, 2018;9(1):4667. 2018 [PMID: 30405100] (Western Blot, Human) Western Blot Human
Z Liu, S Yan, J Wang, Y Xu, Y Wang, S Zhang, X Xu, Q Yang, X Zeng, Y Zhou, X Gu, S Lu, Z Fu, DJ Fulton, NL Weintraub, RB Caldwell, W Zhang, C Wu, XL Liu, JF Chen, A Ahmad, I Kaddour-Dj, M Al-Shabraw, Q Li, X Jiang, Y Sun, A Sodhi, L Smith, M Hong, Y Huo Endothelial adenosine A2a receptor-mediated glycolysis is essential for pathological retinal angiogenesis Nat Commun, 2017;8(1):584. 2017 [PMID: 28928465] (Western Blot, Human) Western Blot Human
L Li, R Zviti, C Ha, ZV Wang, JA Hill, F Lin Forkhead Box O3 (FoxO3) Regulates Kidney Tubular Autophagy Following Urinary Tract Obstruction J. Biol. Chem., 2017;0(0):. 2017 [PMID: 28705935] (Western Blot, Mouse) Western Blot Mouse
X Li, J Lu, Q Kan, X Li, Q Fan, Y Li, R Huang, A Slipicevic, HP Dong, L Eide, J Wang, H Zhang, V Berge, MA Goscinski, G Kvalheim, JM Nesland, Z Suo Metabolic reprogramming is associated with flavopiridol resistance in prostate cancer DU145 cells Sci Rep, 2017;7(1):5081. 2017 [PMID: 28698547] (Western Blot, Human) Western Blot Human
P Himmels, I Paredes, H Adler, A Karakatsan, R Luck, HH Marti, O Ermakova, E Rempel, ET Stoeckli, C Ruiz de Al Motor neurons control blood vessel patterning in the developing spinal cord Nat Commun, 2017;8(0):14583. 2017 [PMID: 28262664] (IHC, Mouse) IHC Mouse
Gabi U Dachs Ascorbate availability affects tumor implantation-take rate and increases tumor rejection in Gulo(-/-) mice Hypoxia (Auckl), 2016;4(0):41-52. 2016 [PMID: 27800507] (Western Blot, Mouse) Western Blot Mouse
Identification of ?-Dystrobrevin as a Direct Target of miR-143: Involvement in Early Stages of Neural Differentiation PLoS ONE, 2016;11(5):e0156325. 2016 [PMID: 27223470] (Western Blot, Human) Western Blot Human
Show All 21 Publications.

Reviews for HIF-1 alpha Antibody (AF1935) (2) 42

Average Rating: 4
(Based on 2 reviews)
We have 2 reviews tested in 2 species: Mouse, mouse brain Tissue and Mouse.

Reviews using AF1935:
Filter by Applications
WB
(1)
All Applications
Filter by Species
Mouse
(1)
mouse brain Tissue and Mouse
(1)
All Species
Images Ratings Applications Species Date Details
 HIF-1 alpha AF1935
Enlarge
4
reviewed by:
Anonymous
mouse brain Tissue and Mouse 10/29/2018
View

Summary

Sample TestedPFA fixed P12 mouse brain Cortex-80µm vibratome,80µm vibratome sections of P12 mouse brain cortex
Speciesmouse brain Tissue and Mouse
LotKAK0214121

Comments

CommentsPerkin Elmer, antigen-antibody signal amplification Kit
  4
reviewed by:
Anonymous
WB Mouse 01/05/2015
View

Summary

ApplicationWestern Blot
Sample TestedSee PMIC 23918831
SpeciesMouse

Blocking

Blocking DetailsSee PMIC 23918831

Details

Detection NotesSee PMIC 23918831

Comments

CommentsPublished: See PMIC 23918831

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for HIF-1 alpha Antibody (AF1935). (Showing 1 - 10 of 11 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, we suggest the following four HIF-1 alpha antibodies based upon customer reviews, as well as the number of peer reviewed publications in which these products have 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 a path exist for detection of HIF 1 in 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. We got the Hif1a (https://www.novusbio.com/products/hif-1-alpha-antibody-h1alpha67_nb100-105 ) antibody from you guys. I used the concentration that is mentioned on your website, but I am getting a band of a completely different size (~70kDa) and not the 120 kDa mentioned.
    • HIF-1 alpha is a notoriously difficult protein to work with due to its rapid degradation. Therefore, the ~70kDa bands are most likely degradation products. It is very important to lyse the cells in hypoxic conditions. We strongly recommend lysing the cells directly into the Laemmli buffer and doing that quickly, so that the exposure to oxygen is minimized.Please go through our hypoxia related FAQs, you should find them very informative:https://www.novusbio.com/support/hypoxia-and-hif-faqsAlso, running a positive control may help confirm the band specificity in your samples. You may prepare them yourself or choose some from our catalog, for example: 1) HeLa Hypoxic / Normoxic Cell Lysate (NBP2-36452)2) HeLa Hypoxic (CoCl2) / Normoxic Cell Lysate (NBP2-36450)
  6. I performed several Western Blots of HIF-1 alpha with different lysis buffers, whole lysates, and cytoplasm/nuclei extractions. I can’t seem to get a good western blot (poor signal, band much lower than expected, etc.). Can someone suggest some technical considerations/tricks I should consider using?
    • A major issue that researchers working with HIF-1 alpha is degradation due to exposure to oxygen. In western blot, this results in a weaker band and/or the appearance of multiple low molecular weight bands (40-80 kDa). We recommend preparing the lysates after collection of cells/tissues as quickly as possible (on ice), preferably in a hypoxic chamber. We also recommend including a true hypoxia mimetic control (eg: cells treated with CoCl2, DMOG… etc.). The controls help distinguish your band of interest from potential degradation/dimer bands.For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: https://www.novusbio.com/support/hypoxia-and-hif-faqs
  7. I am doing HIF1 westerns in HIF-overexpressing mouse liver and adipose tissue using Novus antirabbit HIF1a antibody with overnight incubation. I am getting strong bands around 90kDa. I am aware that HIF theoretical molecular weight is 93kDa, but in westerns, the HIF band is usually around 120kDa according to my internet research. Can someone let me know if I’m getting the right HIF band or just some non-specific bands? Thanks.
    • (1)    HIF-1 alpha’s theoretical molecular weight is 93kDa. The post translationally modified/ubiquitinated form of HIF-1 alpha protein (fails to undergo proteasomal degradation) shows up as a band in the 110-130 kDa range on a Western blot.(2)    The dimeric protein may appear at a position above 200 kDa on non-reducing gels.(3)    Importantly, HIFs are among the most rapidly degradable proteins; therefore, sample preparation is highly important when analyzing HIF1 alpha or HIF2 alpha. When degraded, HIF-1 alpha may show up between 40-80 kDa position on Western blot. Degradation may be avoided by preparing the samples as soon as possible after collection of cells/tissues in hypoxic chamber. Notably, the tissues/cells should be kept on ice during lysate preparation and the lysates should be analyzed as soon as possible.(4)    For troubleshooting suggestions/feedback on more than 25 similar frequently asked questions, I would recommend visiting Novus page: FAQs - Hypoxia and HIFs https://www.novusbio.com/support/hypoxia-and-hif-faqs(5)    Last but not the least, Novus technical support team may be contacted at: technical@novusbio.com 
  8. I have Hif1a nuclear protein extract at -80C. I am wondering if anyone knows how long it would be good for at that temperature since HIf1a is known to be degraded easily.Thank you!
    • You could try a few things to further inhibit the degradation.1) Use the protease inhibitors (if you are not already using them).2) Lyse cells into a buffer that contains SDS or LDS (eg: Laemmli's buffer), since SDS and LDS denature and inhibit proteases. Lysis may even be performed with reducing agents in the buffer (eg. DTT), but this will make your lysates unsuitable for BCA assay.3) Lysing samples rapidly ensures that the samples are instantly homogenized (it also shears DNA released by the SDS).5) Flash-freezing samples in liquid nitrogen rather than freezing at -80*C reduces the window of time for protease activity.6) Freeze samples in individual aliquots, instead of thawing the same vial multiple times.
  9. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
    • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: http://www.novusbio.com/support/hypoxia-and-hif-faqs
  10. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
    • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: https://www.novusbio.com/support/hypoxia-and-hif-faqs
  11. Show All 11 FAQs.

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Blogs on HIF-1 alpha. Showing 1-10 of 33 blog posts - Show all blog posts.

Breast cancer stem cells survive chemotherapy through S100A10-ANXA2-SPT6 interaction that epigenetically promotes OCT4-mediated stemness
By Jamshed Arslan, Pharm D, PhDBreast cancer is the most common cancer among women that causes the greatest number of cancer-related deaths worldwide. After radiotherapy or cytotoxic chemotherapy like paclitax...  Read full blog post.

mTOR Signaling and the Tumor Microenvironment
By Yoskaly Lazo-Fernandez, PhD The mammalian target of rapamycin (mTOR) is a conserved serine/threonine kinase that, as a member of two distinct intracellular protein complexes, mTORC1 and mTORC2, regulates protein...  Read full blog post.

Bad news for stomach cancer: BAMBI protein inhibits gastric carcinoma via TGF-beta/epithelial-mesenchymal transition signaling
By Jamshed Arslan Pharm.D. Gastric carcinoma is the second leading cause of cancer-related deaths worldwide. One of the key features of gastric carcinoma is acidosis, which promotes growth and metastasis of gastric...  Read full blog post.

Developmental regulator Daam2 promotes glial cell tumors by degrading Von Hippel-Lindau protein
By Jamshed Arslan Pharm.D. Glioblastoma is an aggressive type of cancer that forms from the star-shaped glial cells of the central nervous system, called astrocytes. Intriguingly, several genes linked to glioblasto...  Read full blog post.

Stemness for Surviving Hypoxia: TGF-beta/Smad Signaling in Multiple Myeloma
By Jamshed Arslan Pharm.D. Multiple myeloma (MM) is a cancer of antibody-producing plasma cells. The bone marrow (BM) of MM patients is hypoxic, and MM cells overexpress many cancerous genes that are regulated by hy...  Read full blog post.

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.

Showing 1-10 of 33 blog posts - Show all blog posts.
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Recent Reviews

4
5
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4
2
3
0
2
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1
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Anonymous
10/29/2018
Application:
Species: mouse brain Tissue and Mouse

 
Anonymous
01/05/2015
Application: WB
Species: Mouse

Bioinformatics

Gene Symbol HIF1A
Uniprot