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Human CTGF/CCN2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Assay Range
31.3 - 2,000 pg/mL
Please see the product datasheet

* 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 Connective Tissue Growth Factor (CTGF). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma 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)

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CTGF/CCN2

CTGF belongs to the CCN (CYR61/CTGF/NOV) family of secreted proteins that share a common multimodular organization. Each protein contains an IGF-binding protein domain, a von Willebrand factor type C domain, a thrombospondin type I domain, and a cysteine knot domain. The multimodular CTGF has the ability to bind multiple ligands and has numerous effects on development, differentation, and disease. The C-terminal cysteine knot motif contains the heparin and low density lipoprotein receptor (LDLR) binding sites that contribute to the adhesive and mitogenic properties of CTGF.

Long Name:
Connective Tissue Growth Factor
Entrez Gene IDs:
1490 (Human); 14219 (Mouse); 64032 (Rat)
Alternate Names:
CCN2; CCN2IGFBP-8; connective tissue growth factor; CTGF; CTGRP; Fisp12; HCS24; Hypertrophic chondrocyte-specific protein 24; IBP-8; IGF-binding protein 8; IGFBP-8; IGFBP8CCN family member 2; Insulin-like growth factor-binding protein 8; MGC102839; NOV2

Assay Procedure


Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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 CTGF/CCN2 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.

12 Citations: Showing 1 - 10
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  1. The Role of CTGF in Liver Fibrosis Induced in 3D Human Liver Spheroids
    Authors: SR Trampuž, S van Riet, Å Nordling, M Ingelman-S
    Cells, 2023-01-13;12(2):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts
    Authors: KEC Blokland, M Nizamoglu, H Habibie, T Borghuis, M Schuliga, BN Melgert, DA Knight, CA Brandsma, SD Pouwels, JK Burgess
    Frontiers in Pharmacology, 2022-11-03;13(0):989169.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Circular RNA circBNC2 inhibits epithelial cell G2-M arrest to prevent fibrotic maladaptive repair
    Authors: P Wang, Z Huang, Y Peng, H Li, T Lin, Y Zhao, Z Hu, Z Zhou, W Zhou, Y Liu, FF Hou
    Nature Communications, 2022-10-31;13(1):6502.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Airway basal cells show a dedifferentiated KRT17highPhenotype and promote fibrosis in idiopathic pulmonary fibrosis
    Authors: B Jaeger, JC Schupp, L Plappert, O Terwolbeck, N Artysh, G Kayser, P Engelhard, TS Adams, R Zweigerdt, H Kempf, S Lienenklau, W Garrels, I Nazarenko, D Jonigk, M Wygrecka, D Klatt, A Schambach, N Kaminski, A Prasse
    Nature Communications, 2022-09-26;13(1):5637.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Elevated Serotonin and NT-proBNP Levels Predict and Detect Carcinoid Heart Disease in a Large Validation Study
    Authors: S Levy, AB Kilgallen, CM Korse, MIFJ Oerlemans, JPG Sluijter, LW van Laake, GD Valk, MET Tesselaar
    Cancers, 2022-05-10;14(10):.
    Species: Human
    Sample Types: Serum
  6. Interleukin-31 promotes fibrosis and T helper 2 polarization in systemic sclerosis
    Authors: A Kuzumi, A Yoshizaki, KM Matsuda, H Kotani, Y Norimatsu, M Fukayama, S Ebata, T Fukasawa, A Yoshizaki-, Y Asano, K Morikawa, Y Kazoe, K Mawatari, T Kitamori, S Sato
    Nature Communications, 2021-10-12;12(1):5947.
    Species: Human
    Sample Types: Cell Culture Supernates, Cell Lysates, Serum, Tissue Homogenates
  7. Regulation of Cellular Senescence Is Independent from Profibrotic Fibroblast-Deposited ECM
    Authors: KEC Blokland, H Habibie, T Borghuis, GJ Teitsma, M Schuliga, BN Melgert, DA Knight, CA Brandsma, SD Pouwels, JK Burgess
    Cells, 2021-06-29;10(7):.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Pharmacological normalization of pancreatic cancer-associated fibroblast secretome impairs pro-metastatic cross-talk with macrophages: Stromal CSF-1 facilitates metastasis
    Authors: R Samain, A Brunel, T Douché, M Fanjul, S Cassant-So, J Rochotte, J Cros, C Neuzillet, J Raffenne, C Duluc, A Perraud, J Nigri, V Gigoux, I Bieche, M Ponzo, G Carpentier, I Cascone, R Tomasini, HA Schmid, M Mathonnet, R Nicolle, MP Bousquet, Y Martineau, S Pyronnet, C Jean, C Bousquet
    Cellular and Molecular Gastroenterology and Hepatology, 2021-01-20;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Paracrine signalling by cardiac calcitonin controls atrial fibrogenesis and arrhythmia
    Authors: LM Moreira, A Takawale, M Hulsurkar, DA Menassa, A Antanavici, SK Lahiri, N Mehta, N Evans, C Psarros, P Robinson, AJ Sparrow, MA Gillis, N Ashley, P Naud, J Barallobre, K Theofilato, A Lee, M Norris, MV Clarke, PK Russell, B Casadei, S Bhattachar, JD Zajac, RA Davey, M Sirois, A Mead, A Simmons, M Mayr, R Sayeed, G Krasopoulo, C Redwood, KM Channon, JC Tardif, XHT Wehrens, S Nattel, S Reilly
    Nature, 2020-11-04;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. A 3D-Bioprinted dual growth factor-releasing intervertebral disc scaffold induces nucleus pulposus and annulus fibrosus reconstruction
    Authors: B Sun, M Lian, Y Han, X Mo, W Jiang, Z Qiao, K Dai
    Bioact Mater, 2020-08-14;6(1):179-190.
    Species: Rat
    Sample Types: Cell Culture Supernates
  11. Characterization of the T Cell Response to Polypropylene Mesh in Women with Complications
    Authors: L Tennyson, M Rytel, S Palcsey, L Meyn, R Liang, P Moalli
    Am. J. Obstet. Gynecol., 2018-11-09;0(0):.
    Species: Human
    Sample Types: Tissue Homogenates
  12. Mechanism of anti-remodelling action of treprostinil in human pulmonary arterial smooth muscle cells
    Authors: C Lambers, C Kornauth, F Oberndorfe, PM Boehm, M Tamm, W Klepetko, M Roth
    PLoS ONE, 2018-11-01;13(11):e0205195.
    Species: Human
    Sample Types: Cell Culture Supernates


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Reviews for Human CTGF/CCN2 DuoSet ELISA

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By Anonymous on 03/22/2022
Sample Tested: Urine

Poor dilution linearity with urine but also saw a matrix effect with 4-fold urine at the higher end of the standard curve. The matrix effect can most likely be reduced by using 8-fold dilution or higher, but then our samples may be too dilute to measure CTGF concentration.

DuoSets are a little hard to follow because the protocol and reagent dilution information is on three separate pieces of paper.

By Anonymous on 04/07/2020
Sample Tested: Human serum