Human Cyr61/CCN1 Quantikine ELISA Kit

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DCYR10
Control Products Available
Human Cyr61/CCN1 ELISA Standard Curve
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Product Details
Procedure
Citations (2)
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Human Cyr61/CCN1 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Saliva (50 uL), Human Milk (10 uL)
Sensitivity
3.8 pg/mL
Assay Range
39.1 - 2,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Saliva, Human Milk)
Specificity
Natural and recombinant human Cyr61
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human Cyr61/CCN1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Cyr61 in cell culture supernates, serum, plasma, saliva, and human milk. It contains CHO cell-expressed recombinant human Cyr61 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Cyr61 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring Cyr61.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Saliva, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 256 721 1442 249 712 1445
Standard Deviation 5.94 14.2 33.5 15.9 35 73.3
CV% 2.3 2 2.3 6.4 4.9 5.1

Recovery

The recovery of Cyr61 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 107 98-117
EDTA Plasma (n=4) 98 91-104
Serum (n=4) 101 95-113

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Cyr61 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Cyr61/CCN1 ELISA Linearity

Data Examples

Human Cyr61/CCN1 ELISA Standard Curve

Product Datasheets

Preparation and Storage

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

Background: Cyr61/CCN1

Cyr61, also known as IGFBP-10 and CCN1, is a 50 kDa matricellular glycoprotein that regulates the growth and adhesion of vascular endothelial cells, fibroblasts, and monocytes. Cyr61 induces VEGF upregulation, angiogenesis, and tumorigenesis through interactions with integrins alpha V beta 3, alpha V beta 5, alpha M beta 2, and alpha 6 beta 1. Cyr61 is cleaved by plasmin within its VWF domain which generates an N-terminal fragment that is not associated with the matrix but retains the ability to induce endothelial cell migration.

Long Name:
Cysteine-Rich, Angiogenic Inducer 61
Entrez Gene IDs:
3491 (Human); 16007 (Mouse); 83476 (Rat)
Alternate Names:
CCN family member 1; CCN1; CCN1IGF-binding protein 10; Cyr61; Cysteine-rich angiogenic inducer 61; cysteine-rich, angiogenic inducer, 61; GIG1; GIG1cysteine-rich heparin-binding protein 61; IBP-10; IGFBP-10; IGFBP10cysteine-rich, anigogenic inducer, 61; Insulin-like growth factor-binding protein 10; protein CYR61; Protein GIG1
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Cyr61/CCN1 Quantikine ELISA Kit

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.

2 Citations: Showing 1 - 2
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  1. Urinary Cysteine-Rich Protein 61 and Trefoil Factor 3 as Diagnostic Biomarkers for Colorectal Cancer
    Authors: T Shimura, H Iwasaki, M Kitagawa, M Ebi, T Yamada, T Yamada, T Katano, H Nisie, Y Okamoto, K Ozeki, T Mizoshita, H Kataoka
    Transl Oncol, 2019;12(3):539-544.
    Species: Human
    Sample Types: Urine
  2. Combining bevacizumab and chemoradiation in rectal cancer. Translational results of the AXEBeam trial.
    Authors: Verstraete M, Debucquoy A, Dekervel J, Van Pelt J, Verslype C, Devos E, Chiritescu G, Dumon K, D'Hoore A, Gevaert O, Sagaert X, Van Cutsem E, Haustermans K
    Br J Cancer, 2015;112(8):1314-25.
    Species: Human
    Sample Types: Whole Blood

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