Mouse/Rat IGF-I/IGF-1 DuoSet 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 ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse IGF-I. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
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
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 3 (5 plates): (Catalog # DY009) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 3.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY004), or 5% Tween 20 in PBS, 7.2-7.4, 0.2 μm filtered
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: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: IGF-I/IGF-1
Insulin-like Growth Factor 1 (IGF-1), also known as somatomedin C, is a member of the insulin superfamily. It was originally discovered as a mediator of growth hormone actions on somatic cell growth, but has also been shown to be an important regulator of cell metabolism, differentiation and survival. IGF-1 is synthesized as a preproprotein that is proteolytically cleaved to generate the mature protein linked by three disulfide bonds. Mature IGF-1 is highly conserved among large mammals, with 100% sequence identity between the human, bovine, porcine, equine, and canine proteins.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL of Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Mouse/Rat IGF-I/IGF-1 DuoSet 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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Caloric restriction leads to druggable LSD1-dependent cancer stem cells expansion
Authors: Pallavi, R;Gatti, E;Durfort, T;Stendardo, M;Ravasio, R;Leonardi, T;Falvo, P;Duso, BA;Punzi, S;Xieraili, A;Polazzi, A;Verrelli, D;Trastulli, D;Ronzoni, S;Frascolla, S;Perticari, G;Elgendy, M;Varasi, M;Colombo, E;Giorgio, M;Lanfrancone, L;Minucci, S;Mazzarella, L;Pelicci, PG;
Nature communications
Species: Mouse
Sample Types: Cell Culture Supernates
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Long-term effects of a fat-directed FGF21 gene therapy in aged female mice
Authors: Anderson, JM;Arnold, WD;Huang, W;Ray, A;Owendoff, G;Cao, L;
Gene therapy
Species: Mouse
Sample Types: Serum
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mTORC2-NDRG1-CDC42 axis couples fasting to mitochondrial fission
Authors: Martinez-Lopez, N;Mattar, P;Toledo, M;Bains, H;Kalyani, M;Aoun, ML;Sharma, M;McIntire, LBJ;Gunther-Cummins, L;Macaluso, FP;Aguilan, JT;Sidoli, S;Bourdenx, M;Singh, R;
Nature cell biology
Species: Mouse
Sample Types: Serum
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Neural precursor cells tune striatal connectivity through the release of IGFBPL1
Authors: E Butti, S Cattaneo, M Bacigalupp, M Cambiaghi, GM Scotti, E Brambilla, F Ruffini, G Sferruzza, M Ripamonti, F Simeoni, L Cacciaguer, A Zanghì, A Quattrini, R Fesce, P Panina-Bor, F Giannese, D Cittaro, T Kuhlmann, P D'Adamo, MA Rocca, S Taverna, G Martino
Nature Communications, 2022-12-08;13(1):7579.
Species: Mouse
Sample Types: Plasma
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Critical timing of ACEi initiation prevents compensatory glomerular hypertrophy in the remaining single kidney
Authors: AS Naik, SQ Wang, M Chowdhury, J Aqeel, CL O'Connor, JE Wiggins, M Bitzer, RC Wiggins
Scientific Reports, 2021-10-01;11(1):19605.
Species: Rat
Sample Types: Urine
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Recovery of the maternal skeleton after lactation is impaired by advanced maternal age but not by reduced IGF availability in the mouse
Authors: MD Rogowska, UNV Pena, N Binning, JK Christians
PLoS ONE, 2021-09-01;16(9):e0256906.
Species: Mouse
Sample Types: Serum
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CSF1R-dependent macrophages control postnatal somatic growth and organ maturation
Authors: S Keshvari, M Caruso, N Teakle, L Batoon, A Sehgal, OL Patkar, M Ferrari-Ce, CE Snell, C Chen, A Stevenson, FM Davis, SJ Bush, C Pridans, KM Summers, AR Pettit, KM Irvine, DA Hume
PloS Genetics, 2021-06-03;17(6):e1009605.
Species: Rat
Sample Types: Serum
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Magnoliae Cortex Alleviates Muscle Wasting by Modulating M2 Macrophages in a Cisplatin-Induced Sarcopenia Mouse Model
Authors: M Hong, IH Han, I Choi, N Cha, W Kim, SK Kim, H Bae
International Journal of Molecular Sciences, 2021-03-20;22(6):.
Species: Mouse
Sample Types: Tissue Homogenates
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Embryonic stem cell-derived mesenchymal stem cells promote colon epithelial integrity and regeneration by elevating circulating IGF-1 in colitis mice
Authors: J Xu, X Wang, J Chen, S Chen, Z Li, H Liu, Y Bai, F Zhi
Theranostics, 2020-10-30;10(26):12204-12222.
Species: Mouse
Sample Types: Serum
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The TSPO-NOX1 axis controls phagocyte-triggered pathological angiogenesis in the eye
Authors: A Wolf, M Herb, M Schramm, T Langmann
Nat Commun, 2020-06-01;11(1):2709.
Species: Mouse
Sample Types: Tissue Homogenates
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Proteasome Activation by Insulin-like Growth Factor-1/Nuclear Factor Erythroid 2-related Factor 2 Signaling Promotes Exercise-induced Neurogenesis
Authors: X Niu, Y Zhao, N Yang, X Zhao, W Zhang, X Bai, A Li, W Yang, L Lu
Stem Cells, 2019-11-13;0(0):.
Species: Mouse
Sample Types: Serum
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Effects of IGF-1 isoforms on muscle growth and sarcopenia
Authors: F Ascenzi, L Barberi, G Dobrowolny, A Villa Nova, C Nicoletti, E Rizzuto, N Rosenthal, BM Scicchitan, A Musarò
Aging Cell, 2019-04-05;0(0):e12954.
Species: Mouse
Sample Types: Serum
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Inflammation in the hippocampus affects IGF1 receptor signaling and contributes to neurological sequelae in rheumatoid arthritis
Authors: KME Andersson, C Wasén, L Juzokaite, L Leifsdotti, MC Erlandsson, ST Silfverswä, A Stokowska, M Pekna, M Pekny, K Olmarker, RA Heckemann, M Kalm, MI Bokarewa
Proc. Natl. Acad. Sci. U.S.A., 2018-12-03;0(0):.
Species: Mouse
Sample Types: Serum
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Epigenetic Restoration of Fetal-like IGF1 Signaling Inhibits Leukemia Stem Cell Activity
Authors: V Giambra, S Gusscott, D Gracias, R Song, SH Lam, P Panelli, K Tyshchenko, CE Jenkins, C Hoofd, A Lorzadeh, A Carles, M Hirst, CJ Eaves, AP Weng
Cell Stem Cell, 2018-09-27;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Mycobacterium leprae-induced Insulin-like Growth Factor I attenuates antimicrobial mechanisms, promoting bacterial survival in macrophages
Authors: L R Batista-Si
Sci Rep, 2016-06-10;6(0):27632.
Species: Mouse
Sample Types: Cell Culture Supernates
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Hypothalamic gene transfer of BDNF inhibits breast cancer progression and metastasis in middle age obese mice.
Authors: Liu, Xianglan, McMurphy, Travis, Xiao, Run, Slater, Andrew, Huang, Wei, Cao, Lei
Mol Ther, 2014-03-18;22(7):1275-84.
Species: Mouse
Sample Types: Serum
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The lung inflammation and skeletal muscle wasting induced by subchronic cigarette smoke exposure are not altered by a high-fat diet in mice.
Authors: Hansen, Michelle, Chen, Hui, Jones, Jessica, Langenbach, Shenna Y, Vlahos, Ross, Gualano, Rosa C, Morris, Margaret, Anderson, Gary P
PLoS ONE, 2013-11-19;8(11):e80471.
Species: Mouse
Sample Types: Tissue Homogenates
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Interleukin-1 participates in the classical and alternative activation of microglia/macrophages after spinal cord injury.
Authors: Sato A, Ohtaki H, Tsumuraya T, Song D, Ohara K, Asano M, Iwakura Y, Atsumi T, Shioda S
J Neuroinflammation, 2012-04-07;9(0):65.
Species: Mouse
Sample Types: Tissue Homogenates
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Plasma proteome profiles associated with inflammation, angiogenesis, and cancer.
Authors: Kelly-Spratt KS, Pitteri SJ, Gurley KE, Liggitt D, Chin A, Kennedy J, Wong CH, Zhang Q, Buson TB, Wang H, Hanash SM, Kemp CJ
PLoS ONE, 2011-05-12;6(5):e19721.
Species: Mouse
Sample Types: Plasma
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The somatotrope as a metabolic sensor: deletion of leptin receptors causes obesity.
Authors: Childs GV, Akhter N, Haney A
Endocrinology, 2010-11-17;152(1):69-81.
Species: Mouse
Sample Types: Serum
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GRP94 is essential for mesoderm induction and muscle development because it regulates insulin-like growth factor secretion.
Authors: Wanderling S, Simen BB, Ostrovsky O, Ahmed NT, Vogen SM, Gidalevitz T, Argon Y
Mol. Biol. Cell, 2007-07-18;18(10):3764-75.
Species: Mouse
Sample Types: Cell Culture Supernates
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Mutant SOD1(G93A) microglia are more neurotoxic relative to wild-type microglia.
Authors: Xiao Q, Zhao W, Beers DR, Yen AA, Xie W, Henkel JS, Appel SH
J. Neurochem., 2007-06-07;102(6):2008-19.
Species: Mouse
Sample Types: Cell Culture Supernates
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Protective effects of an anti-inflammatory cytokine, interleukin-4, on motoneuron toxicity induced by activated microglia.
Authors: Zhao W, Xie W, Xiao Q, Beers DR, Appel SH
J. Neurochem., 2006-10-02;99(4):1176-87.
Species: Rat
Sample Types: Cell Culture Supernates
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