Human BMP-2 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 BMP-2. 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 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
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 # DY995), or 1% BSA in PBS, pH 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: BMP-2
BMPs are secreted signaling molecules that comprise a subfamily of the TGF-beta superfamily and were originally identified as regulators of cartilage and bone formation. There are at least 20 structurally and functionally related BMPs, most of which play roles in embryogenesis and morphogenesis of various tissues and organs. Biologically active BMPs are usually homodimers containing a characteristic cysteine knot structure. Heterodimers, BMP-2/BMP-7 and BMP-4/BMP-7 have also been suggested to exist and function in vivo. They are more potent inducers of bone formation than their respective homodimers. In addition, heterodimers, but not homodimers, are ventral mesoderm inducers. Heterodimer activity may be mediated by a different or additional receptor subtype.
Decapentaplegic (Dpp) is one of at least five TGF-beta superfamily ligands identified in the Drosophila genome. Dpp, a functional ortholog of mammalian BMP-2 and BMP-4, is a morphogen and plays an essential role in Drosophila development. Dpp regulates embryonic dorsal-ventral polarity and is required for gut morphogenesis and outgrowth and patterning of imaginal disks.
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 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 Human BMP-2 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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Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells
Authors: Donnelly, H;Ross, E;Xiao, Y;Hermantara, R;Taqi, AF;Doherty-Boyd, WS;Cassels, J;Tsimbouri, PM;Dunn, KM;Hay, J;Cheng, A;Meek, RMD;Jain, N;West, C;Wheadon, H;Michie, AM;Peault, B;West, AG;Salmeron-Sanchez, M;Dalby, MJ;
Nature communications
Species: Human
Sample Types: Cell Culture Supernates
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A new mRNA structure prediction based approach to identifying improved signal peptides for bone morphogenetic protein 2
Authors: Wilkinson, P;Jackson, B;Fermor, H;Davies, R;
BMC biotechnology
Species: Human
Sample Types: Cell Culture Supernates
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Bone Regenerative Effect of Injectable Hypoxia Preconditioned Serum-Fibrin (HPS-F) in an Ex Vivo Bone Defect Model
Authors: Jiang, J;Röper, L;Fuchs, F;Hanschen, M;Failer, S;Alageel, S;Cong, X;Dornseifer, U;Schilling, AF;Machens, HG;Moog, P;
International journal of molecular sciences
Species: Human
Sample Types: Serum
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In vitro and in vivo applications of a universal and synthetic thermo-responsive drug delivery hydrogel platform
Authors: H Gholizadeh, E Landh, DM Silva, A Granata, D Traini, P Young, A Fathi, S Maleknia, T Abrams, F Dehghani, H Xin Ong
International journal of pharmaceutics, 2023-02-25;635(0):122777.
Species: N/A
Sample Types: Hydrogel Supernates
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In vitro and in vivo applications of a universal and synthetic thermo-responsive drug delivery hydrogel platform
Authors: H Gholizadeh, E Landh, DM Silva, A Granata, D Traini, P Young, A Fathi, S Maleknia, T Abrams, F Dehghani, H Xin Ong
International journal of pharmaceutics, 2023;635(0):122777.
Species: N/A
Sample Types: Hydrogel Supernates
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Engineering of immunoinstructive extracellular matrices for enhanced osteoinductivity
Authors: A García-Gar, S Pigeot, I Martin
Bioactive materials, 2022-12-21;24(0):174-184.
Species: Human
Sample Types: Tissue Homogenates
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CTR9 drives osteochondral lineage differentiation of human mesenchymal stem cells via epigenetic regulation of BMP-2 signaling
Authors: NT Chan, MS Lee, Y Wang, J Galipeau, WJ Li, W Xu
Science Advances, 2022-11-16;8(46):eadc9222.
Species: Human
Sample Types: Cell Culture Supernates
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Optimization of loading protocols for tissue engineering experiments
Authors: YD Ladner, AR Armiento, EJ Kubosch, JG Snedeker, MJ Stoddart
Scientific Reports, 2022-03-24;12(1):5094.
Species: Human
Sample Types: Cell Culture Supernates
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Efficient healing of large osseous segmental defects using optimized chemically modified messenger RNA encoding BMP-2
Authors: RE De La Vega, M van Griens, W Zhang, MJ Coenen, CV Nagelli, JA Panos, CJ Peniche Si, J Geiger, C Plank, CH Evans, ER Balmayor
Science Advances, 2022-02-16;8(7):eabl6242.
Species: Mouse
Sample Types: Tissue Homogenates
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Gremlin-1 and BMP-4 Overexpressed in Osteoarthritis Drive an Osteochondral-Remodeling Program in Osteoblasts and Hypertrophic Chondrocytes
Authors: ML Pérez-Loza, L Sudre, S van Eegher, D Citadelle, A Pigenet, MH Lafage-Pro, P Pastoureau, F De Ceuninc, F Berenbaum, X Houard
International Journal of Molecular Sciences, 2022-02-14;23(4):.
Species: Human
Sample Types: Cell Culture Supernates
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Engineered bridge protein with dual affinity for bone morphogenetic protein-2 and collagen enhances bone regeneration for spinal fusion
Authors: PS Briquez, HM Tsai, EA Watkins, JA Hubbell
Science Advances, 2021-06-11;7(24):.
Species: Human
Sample Types: Recombinant Protein
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Evaluation of BMP2/miRNA co-expression systems for potent therapeutic efficacy in bone-tissue regeneration
Authors: TK Brenner, K Posa-Marka, D Hercher, S Sperger, P Heimel, C Keibl, S Nürnberger, J Grillari, H Redl, A Hacobian
European cells & materials, 2021-03-03;41(0):245-268.
Species: Hamster, Mouse
Sample Types: Cell Culture Supernates
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Selective endocytosis of recombinant human BMPs through cell surface heparan sulfate proteoglycans in CHO cells: BMP-2 and BMP-7
Authors: MG Kim, CL Kim, YS Kim, JW Jang, GM Lee
Scientific Reports, 2021-02-09;11(1):3378.
Species: Hamster
Sample Types: Cell Culture Supernates
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Zika virus NS3 protease induces bone morphogenetic protein-dependent brain calcification in human fetuses
Authors: W Chen, SS Foo, E Hong, C Wu, WS Lee, SA Lee, D Evseenko, MEL Moreira, A García-Sas, G Cheng, K Nielsen-Sa, P Brasil, E Avvad-Port, JU Jung
Nature Microbiology, 2021-01-28;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Biomimetic Mineralization on 3D Printed PLA Scaffolds: On the Response of Human Primary Osteoblasts Spheroids and In Vivo Implantation
Authors: MOC Maia-Pinto, ACB Brochado, BN Teixeira, SC Sartoretto, MJ Uzeda, ATNN Alves, GG Alves, MD Calasans-M, RMSM Thiré
Polymers, 2020-12-27;13(1):.
Species: Rat
Sample Types: Tissue Homogenates
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TEffects of FK506 on the healing of diaphyseal, critical size defects in the rat femur
Authors: RE De la Vega, MJ Coenen, SA Müller, CV Nagelli, NP Quirk, C Lopez de P, CH Evans
Eur Cell Mater, 2020-10-06;40(0):160-171.
Species: Rat
Sample Types: Cell Culture Supernates
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Detection of circulating BMP5 as a risk factor for Barrett's esophagus
Authors: ACP Correia, S Calpe, N Mostafavi, SJM Hoefnagel, MDC Sancho-Ser, PS de Koning, KK Krishnadat
Scientific Reports, 2020-09-23;10(1):15579.
Species: Human
Sample Types: Serum
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Enhancing the regenerative effectiveness of growth factors by local inhibition of interleukin-1 receptor signaling
Authors: Z Julier, R Karami, B Nayer, YZ Lu, AJ Park, K Maruyama, GA Kuhn, R Müller, S Akira, MM Martino
Sci Adv, 2020-06-12;6(24):eaba7602.
Species: Mouse
Sample Types: Extracellular Matrix
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Enhancing the regenerative effectiveness of growth factors by local inhibition of interleukin-1 receptor signaling
Authors: Z Julier, R Karami, B Nayer, YZ Lu, AJ Park, K Maruyama, GA Kuhn, R Müller, S Akira, MM Martino
Sci Adv, 2020;6(24):eaba7602.
Species: Mouse
Sample Types: Extracellular Matrix
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Synergistic effects of recombinant Lentiviral-mediated BMP2 and TGF-beta3 on the osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro
Authors: W He, L Chen, Y Huang, Z Xu, W Xu, N Ding, J Chen
Cytokine, 2019-04-13;120(0):1-8.
Species: Rat
Sample Types: Cell Culture Supernates
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Large-scale secretome analyses unveil the superior immunosuppressive phenotype of umbilical cord stromal cells as compared to other adult mesenchymal stromal cells
Authors: A Islam, I Urbarova, JA Bruun, I Martinez-Z
Eur Cell Mater, 2019-02-20;37(0):153-174.
Species: Human
Sample Types: Cell Culture Supernates
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TRAIL attenuates RANKL-mediated osteoblastic signalling in vascular cell mono-culture and co-culture models
Authors: E Harper, KD Rochfort, H Forde, C Davenport, D Smith, PM Cummins
PLoS ONE, 2017-11-16;12(11):e0188192.
Species: Human
Sample Types: Cell Culture Supernates
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Contribution of Implanted, Genetically Modified Muscle Progenitor Cells Expressing BMP-2 to New Bone Formation in a Rat Osseous Defect
Authors: RE De La Vega, CL De Padilla, M Trujillo, N Quirk, RM Porter, CH Evans, E Ferreira
Mol. Ther., 2017-10-05;0(0):.
Species: Rat
Sample Types: Tissue Supernates
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A doxycycline inducible, adenoviral BMP-2 gene delivery system to bone
Authors: Jennifer J Bara
J Tissue Eng Regen Med, 2017-06-12;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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GATA4-dependent organ-specific endothelial differentiation controls liver development and embryonic hematopoiesis
Authors: C Géraud, PS Koch, J Zierow, K Klapproth, K Busch, V Olsavszky, T Leibing, A Demory, F Ulbrich, M Diett, S Singh, C Sticht, K Breitkopf-, K Richter, SM Karppinen, T Pihlajanie, B Arnold, HR Rodewald, HG Augustin, K Schledzews, S Goerdt
J. Clin. Invest, 2017-02-20;127(3):1099-1114.
Species: Human
Sample Types: Cell Culture Supernates
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Effect of BMP-2 Delivery Mode on Osteogenic Differentiation of Stem Cells
Authors: T Jung, JH Lee, S Park, YJ Kim, J Seo, HE Shim, KS Kim, HS Jang, HM Chung, SG Oh, SH Moon, SW Kang
Stem Cells Int, 2017-01-19;2017(0):7859184.
Species: Human
Sample Types: Recombinant Protein
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Human cellular CYBA UTR sequences increase mRNA translation without affecting the half-life of recombinant RNA transcripts
Sci Rep, 2016-12-15;6(0):39149.
Species: Mouse
Sample Types: Cell Culture Supernates
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Dual delivery of active antibactericidal agents and bone morphogenetic protein at sustainable high concentrations using biodegradable sheath-core-structured drug-eluting nanofibers
Int J Nanomedicine, 2016-08-17;11(0):3927-37.
Species: Human
Sample Types: Recombinant Protein
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Human Articular Cartilage Progenitor Cells Are Responsive to Mechanical Stimulation and Adenoviral-Mediated Overexpression of Bone-Morphogenetic Protein 2.
Authors: Neumann A, Gardner O, Williams R, Alini M, Archer C, Stoddart M
PLoS ONE, 2015-08-20;10(8):e0136229.
Species: Human
Sample Types: Cell Culture Supernates
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Enhancement of bone regeneration by gene delivery of BMP2/Runx2 bicistronic vector into adipose-derived stromal cells.
Authors: Lee SJ, Kang SW, Do HJ, Han I, Shin DA, Kim JH, Lee SH
Biomaterials, 2010-04-21;31(21):5652-9.
Species: Human
Sample Types: Cell Culture Supernates
FAQs
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Does Human BMP-2 Duoset ELISA (Catalog # DY355 and DY355-05) recognize all forms of BMP-2? Does this kit detect the BMP-2/BMP-7 heterodimer?
Human BMP-2 Duoset (Catalog # DY355 and DY355-05) is specific to mammalian expressed BMP-2. Recombinant E.coli-expressed BMP-2 (Catalog # 355-BEC) is not detected in this assay. Cross-reactivity information is provided under the specificity section of the Duoset insert. The BMP-2/BMP-7 heterodimer shows 11% cross-reactivity in the BMP-2 Duoset ELISA.
Reviews for Human BMP-2 DuoSet ELISA
Average Rating: 4.8 (Based on 4 Reviews)
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