Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA, 2 Plate

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Lysates prepared from NIH-3T3 mouse embryonic fibroblast cells, untreated or treated with 150 µM of CoCl2, were incubated in wells coated with Human/Mouse Total HIF-1 alpha Capture Antibody. Unbound material was ...read more
Lysates were prepared from MCF-7 human breast cancer cells, untreated or treated with 150 µM CoCl2. HIF-1 alpha was measured with the Human/Mouse Total HIF-1 alpha DuoSet IC ELISA and the same lysates were ...read more

Product Details

Summary
Reactivity Hu, MuSpecies Glossary
Applications ELISA
Conjugate
Biotin

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Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA, 2 Plate Summary

Assay Type
Solid Phase Sandwich ELISA
Inter-Assay
See PDF Datasheet for details
Intra-Assay
See PDF Datasheet for details
Spike Recovery
See PDF Datasheet for details
Sample Volume
See PDF Datasheet for details
Gene
HIF1A

Applications/Dilutions

Dilutions
  • ELISA
Application Notes
No significant interference observed with available related molecules.
Publications
Read Publications using DYC1935-2.

Packaging, Storage & Formulations

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Notes

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

Alternate Names for Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA, 2 Plate

  • 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/HIF1A) is the regulated member of the transcription factor heterodimer HIF-1 alpha/beta. HIF-1 alpha/HIF1A binds to hypoxia-response elements (HREs) in the promoters of many genes involved in the adaptation 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 HIF1A.

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Publications for HIF-1 alpha (DYC1935-2)(32)

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


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Human
(20)
Mouse
(10)
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Showing Publications 1 - 10 of 32. Show All 32 Publications.
Publications using DYC1935-2 Applications Species
Shams, S;Stilhano, RS;Silva, EA; Harnessing EGLN1 Gene Editing to Amplify HIF-1 alpha and Enhance Human Angiogenic Response bioRxiv : the preprint server for biology 2023-05-30 [PMID: 37398294] (Human) Human
K P?oszczyca, M Chalimoniu, I Przybylska, M Czuba Effects of Short-Term Phosphate Loading on Aerobic Capacity under Acute Hypoxia in Cyclists: A Randomized, Placebo-Controlled, Crossover Study Nutrients, 2022-01-06;14(2):. 2022-01-06 [PMID: 35057416] (Human) Human
M Jones-Muha, Q Shao, L Cain-Shiel, JP Shaffery, JP Warrington Acid Sensing Ion Channel 2a Is Reduced in the Reduced Uterine Perfusion Pressure Mouse Model and Increases Seizure Susceptibility in Pregnant Mice Cells, 2021-05-08;10(5):. 2021-05-08 [PMID: 34066660] (Mouse) Mouse
J Filtjens, A Roger, L Quatrini, E Wieduwild, J Gouilly, G Hoeffel, R Rossignol, C Daher, G Debroas, S Henri, CM Jones, B Malissen, LK Mackay, A Moqrich, FR Carbone, S Ugolini Nociceptive sensory neurons promote CD8 T cell responses to HSV-1 infection Nature Communications, 2021-05-18;12(1):2936. 2021-05-18 [PMID: 34006861] (Mouse) Mouse
M Okni?ska, B El-Hafny-R, A Paterek, U Mackiewicz, C Crola-Da S, K Brodaczews, M M?czewski, C Kieda Treatment of hypoxia-dependent cardiovascular diseases by myo-inositol trispyrophosphate (ITPP)-enhancement of oxygen delivery by red blood cells J. Cell. Mol. Med., 2020-01-19;0(0):. 2020-01-19 [PMID: 31957267] (Mouse) Mouse
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, 2018-11-13;8(1):35. 2018-11-13 [PMID: 30424786] (Mouse) Mouse
M Li, Y Cui, W He, X Deng, Y Wang, M Cai, Y Wang, J Pei, X Mei, P Wu Effects of Triple-Mutated Hypoxia-Inducible Factor-1? on Angiogenesis and Cardiac Function Improvement in Rats with Myocardial Infarction Cell. Physiol. Biochem., 2018-11-13;50(6):2329-2340. 2018-11-13 [PMID: 30423574] (Rat) Rat
A Sturrock, D Woller, A Freeman, K Sanders, R Paine Consequences of Hypoxia for the Pulmonary Alveolar Epithelial Cell Innate Immune Response J. Immunol., 2018-10-31;0(0):. 2018-10-31 [PMID: 30381478] (Mouse) Mouse
R Terao, M Honjo, M Aihara Apolipoprotein M Inhibits Angiogenic and Inflammatory Response by Sphingosine 1-Phosphate on Retinal Pigment Epithelium Cells Int J Mol Sci, 2017-12-31;19(1):. 2017-12-31 [PMID: 29301231] (Human) Human
Y Guo Role of HIF-1a in regulating autophagic cell survival during cerebral ischemia reperfusion in rats Oncotarget, 2017-10-01;8(58):98482-98494. 2017-10-01 [PMID: 29228704] (Human, Rat) Human, Rat
Show All 32 Publications.

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Product General Protocols

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

FAQs for HIF-1 alpha (DYC1935-2). (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 (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.Also, 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.
  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 (5)    Last but not the least, Novus technical support team may be contacted via email
  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.
  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.
  11. Show All 11 FAQs.

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Blogs on HIF-1 alpha. Showing 1-10 of 37 blog posts - Show all blog posts.

Hypoxia-Dependent CAR Stabilizing Construct in T cells Improves Solid Tumor Targeting and Efficacy
By Victoria Osinski, PhDDespite advances in the development of cancer immunotherapies, those specifically targeting tumors still remains limited. Currently, there is great interest in utilizing chimeric antigen rece...  Read full blog post.

Tired T cells: Hypoxia Drives T cell Exhaustion in the Tumor Microenvironment
By Hunter MartinezThe paradigm shifting view of the immune system being leveraged to target cancer has led to numerous therapeutic breakthroughs. One major cell group responsible for this revelation is a T cell. ...  Read full blog post.

Understanding ‘Y’ in Breast Cancer: Crucial Role of DNA/RNA-binding Protein YB-1 in the Development, Pre-Invasive, and Metastatic Phases
Jamshed Arslan, Pharm D, PhD In the United States, 1 in 8 women will be diagnosed with breast cancer in her lifetime.1 Despite the prevalence, cancer genesis is a mystery. The heterogeneity of cancers makes it diff...  Read full blog post.


  Read full blog post.

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.

Showing 1-10 of 37 blog posts - Show all blog posts.

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Bioinformatics

Gene Symbol HIF1A