Human IGFBP-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 human IGFBP-1. 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.
Normal Goat Serum: (Catalog # DY005)
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)
Normal Goat Serum: (Catalog # DY005)
Scientific Data
Product Datasheets
Preparation and Storage
Background: IGFBP-1
The superfamily of insulin-like growth factor (IGF) binding proteins include the six high-affinity IGF binding proteins (IGFBP) and at least four additional low-affinity binding proteins referred to as IGFBP related proteins (IGFBP-rP). All IGFBP superfamily members are cysteine-rich proteins with conserved cysteine residues, which are clustered in the amino- and carboxy-terminal thirds of the molecule. IGFBPs modulate the biological activities of IGF proteins. Some IGFBPs may also have intrinsic bioactivity that is independent of their ability to bind IGF proteins. Post-translational modifications of IGFBP, including glycosylation, phosphorylation and proteolysis, have been shown to modify the affinities of the binding proteins to IGF. ALS (Acid Labile Subunit) is a liver-derived protein that exists in a ternary complex with Insulin-like Growth Factor (IGF)-binding Protein-3 (IGFBP-3) or IGFBP-5, and either IGF-I or IGF-II. ALS increases the half-life of IGF/IGFBP complexes in circulation.
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 with NGS, 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 Human IGFBP-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.
16
Citations: Showing 1 - 10
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Subtle changes in perivascular endometrial mesenchymal stem cells after local endometrial injury in recurrent implantation failure
Authors: Y Fan, RWK Lee, XW Ng, CE Gargett, JKY Chan
Scientific Reports, 2023-01-05;13(1):225.
Species: Human
Sample Types: Cell Culture Supernates
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Expression and function of the luteinizing hormone choriogonadotropin receptor in human endometrial stromal cells
Authors: ON Mann, CS Kong, ES Lucas, JJ Brosens, AC Hanyaloglu, PJ Brighton
Scientific Reports, 2022-05-21;12(1):8624.
Species: Human
Sample Types: Cell Culture Supernates
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Immune and Inflammatory Proteins in Cord Blood as Predictive Biomarkers of Retinopathy of Prematurity in Preterm Infants
Authors: YJ Park, SJ Woo, YM Kim, S Hong, YE Lee, KH Park
Invest. Ophthalmol. Vis. Sci., 2019-09-03;60(12):3813-3820.
Species: Human
Sample Types: Plasma
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Interconnected Microphysiological Systems for Quantitative Biology and Pharmacology Studies
Authors: CD Edington, WLK Chen, E Geishecker, T Kassis, LR Soenksen, BM Bhushan, D Freake, J Kirschner, C Maass, N Tsamandour, J Valdez, CD Cook, T Parent, S Snyder, J Yu, E Suter, M Shockley, J Velazquez, JJ Velazquez, L Stockdale, JP Papps, I Lee, N Vann, M Gamboa, ME LaBarge, Z Zhong, X Wang, LA Boyer, DA Lauffenbur, RL Carrier, C Communal, SR Tannenbaum, CL Stokes, DJ Hughes, G Rohatgi, DL Trumper, M Cirit, LG Griffith
Sci Rep, 2018-03-14;8(1):4530.
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Decidual regulation of trophoblast is altered in pregnancies at risk of pre-eclampsia
Authors: LB James-Alla, GS Whitley, K Leslie, AE Wallace, JE Cartwright
J. Mol. Endocrinol., 2018-02-07;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
Applications: ELISA Capture -
NLRP7 contributes to in vitro decidualization of endometrial stromal cells
Authors: JY Huang, PH Yu, YC Li, PL Kuo
Reprod. Biol. Endocrinol., 2017-08-15;15(1):66.
Species: Human
Sample Types: Cell Culture Supernates
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Decidualization is Impaired in Endometrial Stromal Cells from Uterine Rudiments in Mayer-Rokitansky-K�ster-Hauser Syndrome
Authors: SY Brucker, S Eisenbeis, J König, M Lamy, MS Salker, N Zeng, H Seeger, M Henes, D Schöller, B Schönfisch, A Staebler, FA Taran, D Wallwiener, K Rall
Cell. Physiol. Biochem, 2017-02-27;41(3):1083-1097.
Species: Human
Sample Types: Cell Culture Supernates
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Higher IGFBP-1 to IGF-1 serum ratio predicts unfavourable survival in patients with nasopharyngeal carcinoma
Authors: X Feng, J Lin, S Xing, W Liu, G Zhang
BMC Cancer, 2017-01-31;17(1):90.
Species: Human
Sample Types: Serum
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Glioblastoma-derived Macrophage Colony-stimulating Factor (MCSF) Induces Microglial Release of Insulin-like Growth Factor-binding Protein 1 (IGFBP1) to Promote Angiogenesis.
Authors: Nijaguna M, Patil V, Urbach S, Shwetha S, Sravani K, Hegde A, Chandramouli B, Arivazhagan A, Marin P, Santosh V, Somasundaram K
J Biol Chem, 2015-08-05;290(38):23401-15.
Species: Human
Sample Types: Cell Culture Supernates
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Heparin and low-molecular-weight heparins modulate the decidualization of human endometrial stromal cells.
Authors: Fluhr H, Spratte J, Ehrhardt J, Steinmuller F, Licht P, Zygmunt M
Fertil. Steril., 2009-12-03;93(8):2581-7.
Species: Human
Sample Types: Cell Culture Supernates
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Manipulating actin dynamics affects human in vitro decidualization.
Authors: Ihnatovych I, Livak M, Reed J, de Lanerolle P, Strakova Z
Biol. Reprod., 2009-04-01;81(1):222-30.
Species: Human
Sample Types: Cell Culture Supernates
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The androgen and progesterone receptors regulate distinct gene networks and cellular functions in decidualizing endometrium.
Authors: Cloke B, Huhtinen K, Fusi L, Kajihara T, Yliheikkila M, Ho KK, Teklenburg G, Lavery S, Jones MC, Trew G, Kim JJ, Lam EW, Cartwright JE, Poutanen M, Brosens JJ
Endocrinology, 2008-05-29;149(9):4462-74.
Species: Human
Sample Types: Cell Culture Supernates
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Insulin-like growth factor-II (IGF-II), IGF-binding protein-3 (IGFBP-3), and IGFBP-4 in follicular fluid are associated with oocyte maturation and embryo development.
Authors: Wang TH, Chang CL, Wu HM, Chiu YM, Chen CK, Wang HS
Fertil. Steril., 2006-11-01;86(5):1392-401.
Species: Human
Sample Types: Follicular Fluid
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Stromal cells from endometriotic lesions and endometrium from women with endometriosis have reduced decidualization capacity.
Authors: Klemmt PA, Carver JG, Kennedy SH, Koninckx PR, Mardon HJ
Fertil. Steril., 2006-03-01;85(3):564-72.
Species: Human
Sample Types: Cell Culture Supernates
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Genetic regulation of the variation of circulating insulin-like growth factors and leptin in human pedigrees.
Authors: Pantsulaia Ia, Pantsulaia I, Trofimov S, Kobyliansky E, Livshits G
Metab. Clin. Exp., 2005-07-01;54(7):975-81.
Species: Human
Sample Types: Plasma
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Progesterone inhibits insulin-like growth factor binding protein-1 (IGFBP-1) production by explants of the Fallopian tube.
Authors: Davies S, Richardson MC, Anthony FW, Mukhtar D, Cameron IT
Mol. Hum. Reprod., 2004-10-22;10(12):935-9.
Species: Human
Sample Types: Cell Culture Supernates
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Excellent results, very tight replicates on a 384-well format. Sample diluted 1/100 for better resolution