Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA Summary
* 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
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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.
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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
Preparation and Storage
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.
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.
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Citations: Showing 1 - 10
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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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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.
