Human PDGF-BB 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 PDGF-BB. 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: PDGF-BB
The platelet-derived growth factor (PDGF) family of disulfide-linked dimeric proteins consists of four homodimeric proteins, PDGF-AA, PDGF-BB, PDGF-CC and PDGF-DD, and one heterodimeric protein, PDGF-AB. Members of this protein family act mainly on connective tissue. The dimeric isoforms of PDGF are expressed by different cell types and at different times during embryonic development. All PDGF isoforms are synthesized as inactive precursors and are processed to active forms that contain a PDGF/VEGF cysteine knot domain. Only PDGF-C and PDGF-D, also named spinal cord-derived growth factors SCDGF and SCDGF-B, also contain an N-terminal CUB domain. Mature PDGF dimers bind and induce the homo- or hetero-dimerization of two receptor tyrosine kinases (PDGF R alpha and R beta).
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 PDGF-BB 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.
27
Citations: Showing 1 - 10
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Baseline circulating biomarkers, their changes, and subsequent suicidal ideation and depression severity at 6 months: A prospective analysis in patients with mood disorders
Authors: Lengvenyte, A;Cognasse, F;Hamzeh-Cognasse, H;Sénèque, M;Strumila, R;Olié, E;Courtet, P;
Psychoneuroendocrinology
Species: Human
Sample Types: Plasma
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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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Characterization of platelet rich plasma in feline immunodeficiency virus-infected cats: Cell, and PDGF-BB and TGF-beta 1 growth factor analysis
Authors: Miguel-Pastor, L;Satué, K;Chicharro, D;Damiá, E;Cuervo, B;Torres-Torrillas, M;Martins, E;Velasco-Martínez, MG;Carrillo, JM;Sopena, JJ;Cerón, JJ;Rubio, M;
Research in veterinary science
Species: Feline
Sample Types: Platelet-Poor Plasma, Platelet-Rich Plasma
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Hypoxia Preconditioned Serum (HPS) Promotes Proliferation and Chondrogenic Phenotype of Chondrocytes In Vitro
Authors: Jiang, J;Altammar, J;Cong, X;Ramsauer, L;Steinbacher, V;Dornseifer, U;Schilling, AF;Machens, HG;Moog, P;
International journal of molecular sciences
Species: Human
Sample Types: Serum
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Effectiveness of BMP-2 and PDGF-BB Adsorption onto a Collagen/Collagen-Magnesium-Hydroxyapatite Scaffold in Weight-Bearing and Non-Weight-Bearing Osteochondral Defect Bone Repair: In Vitro, Ex Vivo and In Vivo Evaluation
Authors: J Xu, S Fahmy-Garc, MA Wesdorp, N Kops, L Forte, C De Luca, MM Misciagna, L Dolcini, G Filardo, M Labberté, K Vancíková, J Kok, B van Rietbe, J Nickel, E Farrell, PAJ Brama, GJVM van Osch
Journal of functional biomaterials, 2023-02-16;14(2):.
Species: Human
Sample Types: Complex Sample Type
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In Vitro Comparison of Lymphangiogenic Potential of Hypoxia Preconditioned Serum (HPS) and Platelet-Rich Plasma (PRP)
Authors: J Jiang, X Cong, S Alageel, U Dornseifer, AF Schilling, E Hadjipanay, HG Machens, P Moog
International Journal of Molecular Sciences, 2023-01-19;24(3):.
Species: Human
Sample Types: Hypoxia Preconditioned Serum
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In situ regeneration of nasal septal defects using acellular cartilage enhanced with platelet-derived growth factor
Authors: H Lena, G David, K Johann, J Yvonne, Z Frank G, H Daniela, B Roman, B Rolf E, R Nicole
Journal of tissue engineering, 2022-09-20;13(0):2041731422111.
Species: Porcine
Sample Types: Cell Culture Supernates
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Growth Factor Release within Liquid and Solid PRF
Authors: K Zwittnig, B Kirnbauer, N Jakse, P Schlenke, I Mischak, S Ghanaati, S Al-Maawi, D Végh, M Payer, TA Zrnc
Journal of Clinical Medicine, 2022-08-29;11(17):.
Species: Human
Sample Types: Serum
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Therapeutic Effects of Platelet-Derived Extracellular Vesicles in a Bioengineered Tendon Disease Model
Authors: AL Graça, RMA Domingues, I Calejo, M Gómez-Flor, ME Gomes
International Journal of Molecular Sciences, 2022-03-09;23(6):.
Species: Human
Sample Types: Extracellular Vesicles
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Circulating Interleukin-6 and CD16 positive monocytes increase following angioplasty of an arteriovenous fistula
Authors: S Hakki, EJ Robinson, MG Robson
Scientific Reports, 2022-01-26;12(1):1427.
Species: Human
Sample Types: Plasma
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Engagement of the CXCL12-CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis
Authors: SF Mause, E Ritzel, A Deck, F Vogt, EA Liehn
International Journal of Molecular Sciences, 2022-01-14;23(2):.
Species: Human
Sample Types: Cell Culture Supernates
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Intramammary treatment using allogeneic pure platelet-rich plasma in cows with subclinical mastitis caused by Gram-positive bacteria
Authors: PC Duque-Madr, J Velasco-Bo, A Ceballos-M, C López, JU Carmona
Scientific Reports, 2021-12-09;11(1):23737.
Species: Bovine
Sample Types: Plasma
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Why PRP works only on certain patients with tennis elbow? Is PDGFB gene a key for PRP therapy effectiveness? A prospective cohort study
Authors: P Niemiec, K Szyluk, A Balcerzyk, M Kalita, A Jarosz, J Iwanicka, T Iwanicki, T Nowak, M Negru, T Francuz, W Garczorz, W Grzeszczak, S Górczy?ska, W Kania, I ?ak
Bmc Musculoskeletal Disorders, 2021-08-18;22(1):710.
Species: Human
Sample Types: Plasma
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Regional hyperthermia enhances mesenchymal stem cell recruitment to tumor stroma: Implications for mesenchymal stem cell-based tumor therapy
Authors: M Tutter, C Schug, KA Schmohl, S Urnauer, C Kitzberger, N Schwenk, M Petrini, C Zach, S Ziegler, P Bartenstei, WA Weber, G Multhoff, E Wagner, LH Lindner, PJ Nelson, C Spitzweg
Mol Ther, 2020-10-15;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Standing wave-assisted acoustic droplet vaporization for single and dual payload release in acoustically-responsive scaffolds
Authors: M Aliabouzar, A Jivani, X Lu, OD Kripfgans, JB Fowlkes, ML Fabiilli
Ultrason Sonochem, 2020-03-26;66(0):105109.
Species: N/A
Sample Types: Complex Sample Type
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Ultrasonic Generation of Pulsatile and Sequential Therapeutic Delivery Profiles from Calcium-Crosslinked Alginate Hydrogels
Authors: T Emi, K Michaud, E Orton, G Santilli, C Linh, M O'Connell, F Issa, S Kennedy
Molecules, 2019-03-16;24(6):.
Species: Human
Sample Types: Hydrogel Fluid
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Long-term cytokine and growth factor release from equine platelet-rich fibrin clots obtained with two different centrifugation protocols
Authors: RF Jiménez-Ar, C López, ME Álvarez, C Giraldo, M Prades, JU Carmona
Cytokine, 2017-06-23;97(0):149-155.
Species: Equine
Sample Types: Plasma
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The PDGF-BB-SOX7 axis-modulated IL-33 in pericytes and stromal cells promotes metastasis through tumour-associated macrophages
Nat Commun, 2016-05-06;7(0):11385.
Species: Human
Sample Types: Cell Lysates
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Regulating VEGF signaling in platelet concentrates via specific VEGF sequestering
Authors: DG Belair, NN Le, WL Murphy
Biomater Sci, 2016-03-24;4(5):819-25.
Species: Human
Sample Types: Plasma
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Effects of sodium citrate and acid citrate dextrose solutions on cell counts and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel.
Authors: Giraldo C, Alvarez M, Carmona J
BMC Vet Res, 2015-03-14;11(0):60.
Species: Equine
Sample Types: Plasma
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Effects of the breed, sex and age on cellular content and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel.
Authors: Giraldo C, Lopez C, Alvarez M, Samudio I, Prades M, Carmona J
BMC Vet Res, 2013-02-12;9(0):29.
Species: Equine
Sample Types: Plasma
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The 12th-14th type III repeats of fibronectin function as a highly promiscuous growth factor-binding domain.
Authors: Martino MM, Hubbell JA
FASEB J., 2010-07-29;24(12):4711-21.
Species: Human
Sample Types: Protein
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Improvement in disability after alemtuzumab treatment of multiple sclerosis is associated with neuroprotective autoimmunity.
Authors: Jones JL, Anderson JM, Phuah CL, Fox EJ, Selmaj K, Margolin D, Lake SL, Palmer J, Thompson SJ, Wilkins A, Webber DJ, Compston DA, Coles AJ
Brain, 2010-07-21;133(0):2232-47.
Species: Human
Sample Types: Cell Culture Supernates
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Normal ranges of angiogenesis regulatory proteins in human platelets.
Authors: Peterson JE, Zurakowski D, Italiano JE
Am. J. Hematol., 2010-07-01;85(7):487-93.
Species: Human
Sample Types: Cell Lysates
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Soluble forms of VEGF receptor-1 and -2 promote vascular maturation via mural cell recruitment.
Authors: Lorquet S, Berndt S, Blacher S, Gengoux E, Peulen O, Maquoi E, Noel A, Foidart JM, Munaut C, Pequeux C
FASEB J., 2010-05-19;24(10):3782-95.
Species: Human
Sample Types: Cell Culture Supernates
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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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Vascular endothelial growth factor can signal through platelet-derived growth factor receptors.
Authors: Ball SG, Shuttleworth CA, Kielty CM
J. Cell Biol., 2007-04-30;177(3):489-500.
Species: Human
Sample Types: Cell Culture Supernates
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