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

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Standard Curve
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Product Details
Citations (32)
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Reviews (6)

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

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Assay Length
4 hours 40 mins (after plate preparation)
Sample Volume Required
Cell lysates (100 µL)
Assay Range
125.0 - 8,000 pg/mL
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total HIF-1 alpha in cell lysates. An immobilized capture antibody specific for HIF-1 alpha binds both phosphorylated and unphosphorylated HIF-1 alpha. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Available in 2, 5, and 15- (96-well) plate pack sizes
  • Economical alternative to Western blot

Kit Content

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control
  • Streptavidin-HRP

Other Reagents Required

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product 

Scientific Data

Standard Curve

Specificity of Human/Mouse HIF-1 alpha DuoSet IC ELISA is shown by Western Blot. 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 removed by washing, and bound material was solubilized in SDS gel sample buffer. Captured proteins were electrophoresed, transferred to PVDF membranes and immunoblotted with the Human/Mouse Total HIF-1 alpha Detection Antibody.

Amounts of HIF-1 alpha, as Quantified by the Human/Mouse Total HIF-1 alpha DuoSet IC ELISA, are Consistent with the Relative Amounts of HIF-1 alpha Determined by Qualitative Western Blot Analysis. 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 immunoblotted (inset) with Goat Anti-HIF-1 alpha Antibody (Catalog # AF1935). The DuoSet IC ELISA results correlate with the relative amounts of HIF-1 alpha detected by Western blot.

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: HIF-1 alpha/HIF1A

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.

Long Name:
Hypoxia Inducible Factor 1 Subunit Alpha
Entrez Gene IDs:
3091 (Human); 15251 (Mouse); 29560 (Rat)
Alternate Names:
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

Citations for Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

32 Citations: Showing 1 - 10
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  1. Harnessing EGLN1 Gene Editing to Amplify HIF-1 alpha and Enhance Human Angiogenic Response
    Authors: Shams, S;Stilhano, RS;Silva, EA;
    bioRxiv : the preprint server for biology
    Species: Human
    Sample Types: Cell Lysates
  2. Effects of Short-Term Phosphate Loading on Aerobic Capacity under Acute Hypoxia in Cyclists: A Randomized, Placebo-Controlled, Crossover Study
    Authors: K P?oszczyca, M Chalimoniu, I Przybylska, M Czuba
    Nutrients, 2022-01-06;14(2):.
    Species: Human
    Sample Types: Plasma
  3. Nociceptive sensory neurons promote CD8 T cell responses to HSV-1 infection
    Authors: 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
    Nature Communications, 2021-05-18;12(1):2936.
    Species: Mouse
    Sample Types: Tissue Homogenates
  4. Acid Sensing Ion Channel 2a Is Reduced in the Reduced Uterine Perfusion Pressure Mouse Model and Increases Seizure Susceptibility in Pregnant Mice
    Authors: M Jones-Muha, Q Shao, L Cain-Shiel, JP Shaffery, JP Warrington
    Cells, 2021-05-08;10(5):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  5. Treatment of hypoxia-dependent cardiovascular diseases by myo-inositol trispyrophosphate (ITPP)-enhancement of oxygen delivery by red blood cells
    Authors: M Okni?ska, B El-Hafny-R, A Paterek, U Mackiewicz, C Crola-Da S, K Brodaczews, M M?czewski, C Kieda
    J. Cell. Mol. Med., 2020-01-19;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
  6. HIF prolyl hydroxylase inhibition protects skeletal muscle from eccentric contraction-induced injury
    Authors: 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
    Skelet Muscle, 2018-11-13;8(1):35.
    Species: Mouse
    Sample Types: Cell Lysates
  7. Effects of Triple-Mutated Hypoxia-Inducible Factor-1? on Angiogenesis and Cardiac Function Improvement in Rats with Myocardial Infarction
    Authors: M Li, Y Cui, W He, X Deng, Y Wang, M Cai, Y Wang, J Pei, X Mei, P Wu
    Cell. Physiol. Biochem., 2018-11-13;50(6):2329-2340.
    Species: Rat
    Sample Types: Tissue Homogenates
  8. Consequences of Hypoxia for the Pulmonary Alveolar Epithelial Cell Innate Immune Response
    Authors: A Sturrock, D Woller, A Freeman, K Sanders, R Paine
    J. Immunol., 2018-10-31;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
  9. Apolipoprotein M Inhibits Angiogenic and Inflammatory Response by Sphingosine 1-Phosphate on Retinal Pigment Epithelium Cells
    Authors: R Terao, M Honjo, M Aihara
    Int J Mol Sci, 2017-12-31;19(1):.
    Species: Human
    Sample Types: Cell Lysates
  10. Iron-Chelating Drugs Enhance Cone Photoreceptor Survival in a Mouse Model of Retinitis Pigmentosa
    Authors: K Wang, B Peng, J Xiao, O Weinreb, MBH Youdim, B Lin
    Invest. Ophthalmol. Vis. Sci., 2017-10-01;58(12):5287-5297.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. Role of HIF-1a in regulating autophagic cell survival during cerebral ischemia reperfusion in rats
    Authors: Y Guo
    Oncotarget, 2017-10-01;8(58):98482-98494.
    Species: Human, Rat
    Sample Types: Cell Lysates, Tissue Homogenates
  12. The Proangiogenic Capabilities of Malignant Ascites Generated by Aggressive Ovarian Tumors
    Authors: J Miku?a-Pie, P Uruski, S Szubert, K Maksin, R Moszy?ski, D Szpurek, A Wo?niak, S Sajdak, A Tykarski, K Ksi??ek
    Biomed Res Int, 2017-09-20;2017(0):2592496.
    Species: Human
    Sample Types: Cell Lysates
  13. Effects of Histone Deacetylase Inhibitor (Valproic Acid) on the Expression of Hypoxia-inducible Factor-1 Alpha in Human Retinal M�ller Cells
    Authors: YJ Kim, SJ Park, NR Kim, HS Chin
    Korean J Ophthalmol, 2017-02-02;31(1):80-85.
    Species: Human
    Sample Types: Cell Lysates
  14. VEGF released by deferoxamine preconditioned mesenchymal stem cells seeded on collagen-GAG substrates enhances neovascularization
    Sci Rep, 2016-11-10;6(0):36879.
    Species: Human
    Sample Types: Cell Lysates
  15. Glycolysis determines dichotomous regulation of T cell subsets in hypoxia
    Authors: Yang Xu
    J Clin Invest, 2016-06-13;0(0):.
    Species: Human
    Sample Types: Whole Cells
  16. Synchronous down-modulation of miR-17 family members is an early causative event in the retinal angiogenic switch.
    Authors: Nunes D, Dias-Neto E, Cardo-Vila M, Edwards J, Dobroff A, Giordano R, Mandelin J, Brentani H, Hasselgren C, Yao V, Marchio S, Pereira C, Passetti F, Calin G, Sidman R, Arap W, Pasqualini R
    Proc Natl Acad Sci U S A, 2015-03-09;112(12):3770-5.
    Species: Human
    Sample Types: Cell Lysates
  17. Lysyl hydroxylase 2 induces a collagen cross-link switch in tumor stroma.
    Authors: Chen, Yulong, Terajima, Masahiko, Yang, Yanan, Sun, Li, Ahn, Young-Ho, Pankova, Daniela, Puperi, Daniel S, Watanabe, Takeshi, Kim, Min P, Blackmon, Shanda H, Rodriguez, Jaime, Liu, Hui, Behrens, Carmen, Wistuba, Ignacio, Minelli, Rosalba, Scott, Kenneth, Sanchez-Adams, Johannah, Guilak, Farshid, Pati, Debanand, Thilaganathan, Nishan, Burns, Alan R, Creighton, Chad J, Martinez, Elisabet, Zal, Tomasz, Grande-Allen, K Jane, Yamauchi, Mitsuo, Kurie, Jonathan
    J Clin Invest, 2015-02-09;125(3):1147-62.
    Species: Human
    Sample Types: Cell Lysates
  18. HIF1alpha is an independent prognostic factor for overall survival in advanced primary epithelial ovarian cancer - a study of the OVCAD Consortium.
    Authors: Braicu E, Luketina H, Richter R, Cacsire Castillo-Tong D, Lambrechts S, Mahner S, Concin N, Mentze M, Zeillinger R, Vergote I, Sehouli J
    Onco Targets Ther, 2014-09-11;7(0):1563-9.
    Species: Human
    Sample Types: Whole Tissue
  19. Exogenous sphingosine-1-phosphate boosts acclimatization in rats exposed to acute hypobaric hypoxia: assessment of haematological and metabolic effects.
    Authors: Chawla S, Rahar B, Singh M, Bansal A, Saraswat D, Saxena S
    PLoS ONE, 2014-06-02;9(6):e98025.
    Species: Rat
    Sample Types: Tissue Homogenates
  20. Autocrine impact of VEGF-A on uveal melanoma cells.
    Authors: Koch K, Refaian N, Hos D, Schlereth S, Bosch J, Cursiefen C, Heindl L
    Invest Ophthalmol Vis Sci, 2014-04-25;55(4):2697-704.
    Species: Human
    Sample Types: Cell Lysates
  21. GABARBP down-regulates HIF-1alpha expression through the VEGFR-2 and PI3K/mTOR/4E-BP1 pathways.
    Authors: Park S, Kim B, Lee J, Park S, Lee S, Dong S, Rho S
    Cell Signal, 2014-03-29;26(7):1506-13.
    Species: Human
    Sample Types: Nuclear Extract
  22. Identification of the angiogenic gene signature induced by EGF and hypoxia in colorectal cancer.
    Authors: Khong T, Thairu N, Larsen H, Dawson P, Kiriakidis S, Paleolog E
    BMC Cancer, 2013-11-02;13(0):518.
    Species: Human
    Sample Types: Cell Lysates
  23. Regulation of hypoxia-inducible factor 1alpha (HIF-1alpha) by lysophosphatidic acid is dependent on interplay between p53 and Kruppel-like factor 5.
    Authors: Lee S, No Y, Dang D, Dang L, Yang V, Shim H, Yun C
    J Biol Chem, 2013-07-23;288(35):25244-53.
    Species: Human
    Sample Types: Cell Lysates
  24. Hypoxia and oxidative stress markers in pediatric patients undergoing hemodialysis: cross section study.
    Authors: Hamed E, El-Abaseri T, Mohamed A, Ahmed A, El-Metwally T
    BMC Nephrol, 2012-10-13;13(0):136.
    Species: Human
    Sample Types: Plasma
  25. Von Hippel-Lindau protein in the RPE is essential for normal ocular growth and vascular development.
    Authors: Lange CA, Luhmann UF, Mowat FM
    Development, 2012-05-23;139(13):2340-50.
    Species: Mouse
    Sample Types: Tissue Homogenates
  26. Overexpression of carbonic anhydrase and HIF-1alpha in Wilms tumours.
    Authors: Dungwa JV, Hunt LP, Ramani P
    BMC Cancer, 2011-09-12;11(0):390.
    Species: Human
    Sample Types: Tissue Homogenates
  27. Hydroxylase inhibition abrogates TNF-alpha-induced intestinal epithelial damage by hypoxia-inducible factor-1-dependent repression of FADD.
    Authors: Hindryckx P, De Vos M, Jacques P, Ferdinande L, Peeters H, Olievier K, Bogaert S, Brinkman B, Vandenabeele P, Elewaut D, Laukens D
    J. Immunol., 2010-10-13;185(10):6306-16.
    Species: Mouse
    Sample Types: Cell Lysates
  28. Epidermal growth factor receptor regulates MET levels and invasiveness through hypoxia-inducible factor-1alpha in non-small cell lung cancer cells.
    Authors: Xu L, Nilsson MB, Saintigny P, Cascone T, Herynk MH, Du Z, Nikolinakos PG, Yang Y, Prudkin L, Liu D, Lee JJ, Johnson FM, Wong KK, Girard L, Gazdar AF, Minna JD, Kurie JM, Wistuba II, Heymach JV
    Oncogene, 2010-02-15;29(18):2616-27.
    Species: Human
    Sample Types: Cell Lysates
  29. Specific inhibition of hypoxia inducible factor 1 exaggerates cell injury induced by in vitro ischemia through deteriorating cellular redox environment.
    Authors: Guo S, Miyake M, Liu KJ, Shi H
    J. Neurochem., 2009-01-29;108(5):1309-21.
    Species: Human
    Sample Types: Cell Lysates
  30. Role of vitamin D receptor in the antiproliferative effects of calcitriol in tumor-derived endothelial cells and tumor angiogenesis in vivo.
    Authors: Chung I, Han G, Seshadri M, Gillard BM, Yu WD, Foster BA, Trump DL, Johnson CS
    Cancer Res., 2009-01-13;69(3):967-75.
    Species: Mouse
    Sample Types: Cell Lysates
  31. Hypoxia induces leptin gene expression and secretion in human preadipocytes: differential effects of hypoxia on adipokine expression by preadipocytes.
    Authors: Wang B, Wood IS, Trayhurn P
    J. Endocrinol., 2008-05-07;198(1):127-34.
    Species: Human
    Sample Types: Cell Lysates
  32. The aryl hydrocarbon receptor (AhR) inhibits vanadate-induced vascular endothelial growth factor (VEGF) production in TRAMP prostates.
    Authors: Fritz WA, Lin TM, Peterson RE
    Carcinogenesis, 2008-03-20;29(5):1077-82.
    Species: Mouse
    Sample Types: Tissue Homogenates


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Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
By Anonymous on 05/22/2023
Sample Tested: EDTA Plasma

Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
By Anonymous on 05/17/2023
Sample Tested: cell supernatant

Human/Mouse Total HIF-1 alpha DuoSet IC ELISA
By Anonymous on 08/01/2019
Sample Tested: Tumor cell lyastes

Whole tumor lysate was acquired and protein quantified using BCA. 100 ug protein was added to each well from each sample. ELISA was run as per protocol.

Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
By Anonymous on 03/12/2019
Sample Tested: Serum

Human/Mouse Total HIF-1 alpha DuoSet IC ELISA
By Sophie Millar on 01/16/2019
Sample Tested: Cell culture supernatant

Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
By Anonymous on 11/10/2018
Sample Tested: Plasma