ELISA Kit for Connective Tissue Growth Factor (CTGF)

CCN2; HCS24; IGFBP8; NOV2; Insulin-Like Growth Factor-Binding Protein 8; Hypertrophic Chondrocyte-Specific Protein 24; CCN Family Member 2

Specificity

This assay has high sensitivity and excellent specificity for detection of Connective Tissue Growth Factor (CTGF).
No significant cross-reactivity or interference between Connective Tissue Growth Factor (CTGF) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Connective Tissue Growth Factor (CTGF) and the recovery rates were calculated by comparing the measured value to the expected amount of Connective Tissue Growth Factor (CTGF) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-95 88
EDTA plasma(n=5) 89-104 95
heparin plasma(n=5) 78-89 82

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Connective Tissue Growth Factor (CTGF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Connective Tissue Growth Factor (CTGF) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Connective Tissue Growth Factor (CTGF) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 84-105% 81-91% 88-95% 81-94%
EDTA plasma(n=5) 92-99% 78-93% 97-105% 94-105%
heparin plasma(n=5) 79-93% 84-93% 78-96% 98-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Journal of Orthopaedic Research Promotion of bone formation by naringin in a titanium particle-induced diabetic murine calvarial osteolysis model PubMed: 19824055
World Journal of Gastroenterology Connective tissue growth factor hammerhead ribozyme attenuates human hepatic stellate cell function PubMed: PMC2726461
PLos One Activation of JNK Signaling Mediates Connective Tissue Growth Factor Expression and Scar Formation in Corneal Wound Healing Plosone: 0032128
Clinical Biochemistry Clinical significance of inflammatory and fibrogenic cytokines in diabetic nephropathy ScienceDirect: S0009912012000987
The Journal of Heart and Lung Transplantation Connective tissue growth factor (CTGF/CCN2): A protagonist in cardiac allograft vasculopathy development? PubMed: S1053249812010121
Biomedical and Environmental Sciences The Intervention Effect of Rosiglitozone in Ovarian Fibrosis of PCOS Rats Besjournal: Source
Disease Markers Concentrations of connective tissue growth factor in patients with nonalcoholic fatty liver disease: Association with liver fibrosis PubMed: 22846210
World Journal of Gastroenterology Clinical significance of connective tissue growth factor in hepatitis B virus-induced hepatic fibrosis PubMed: PMC3351780
Journal of ethnopharmacology Multitargeted protective effect of Abacopteris penangiana against carrageenan-induced chronic prostatitis in rats. Pubmed: 24211397
Radiation Oncology Cytokine levels as biomarkers of radiation fibrosis in patients treated with breast radiotherapy Ro-journal: Source
PLoS One. Knock Out of S1P3 Receptor Signaling Attenuates Inflammation and Fibrosis in Bleomycin-Induced Lung Injury Mice Model Pubmed:Pmc4157792
European Journal of Heart Failure Connective tissue growth factor and cardiac diastolic dysfunction: human data from the Taiwan Diastolic Heart Failure Registry and molecular basis by cellular and animal models Pubmed:24464932
Bone Connective Tissue Growth Factor is a Target of Notch Signaling in Cells of the Osteoblastic Lineage Pubmed:24792956
American Journal of Pathology Disruption of collagen homeostasis can reverse established age-related myocardial fibrosis Pubmed:25701883
J Gastroenterol Beneficial effects of combined ursodeoxycholic acid and angiotensin-II type 1 receptor blocker on hepatic fibrogenesis in a rat model of nonalcoholic steatohepatitis PubMed: 26190501
Life Sciences Suppression of connective tissue growth factor mediates the renoprotective effect of Sitagliptin rather than Pioglitazone in type 2 diabetes mellitus Pubmed:27049870
International Journal of Molecular Sciences TGF-β and Physiological Root Resorption of Deciduous Teeth. pubmed:28035998
Inflammation Research Fibrosis of extracellular matrix is related to the duration of the disease but is unrelated to the dynamics of collagen metabolism in dilated cardiomyopathy pubmed:27516211
Heart & Vessels Left ventricular reverse remodeling is not related to biopsy-detected extracellular matrix fibrosis and serum markers of fibrosis in dilated cardiomyopathy, regardless of the definition used for LVRR pubmed:28004175
British Journal of Ophthalmology The relationship between anti-vascular endothelial growth factor and fibrosis in proliferative retinopathy: clinical and laboratory evidence pubmed:27531356
Pathophysiology Molecular mechanisms underlying fibrosis and elastin destruction in childhood interstitial lung diseases pubmed:27686729
Evidence-Based Complementary and Alternative Medicine Cardioprotection against Heart Failure by Shenfu Injection via TGF-β/Smads Signaling Pathway pubmed:28698735
Heart and Vessels Left ventricular reverse remodeling is not related to biopsy‑detected extracellular matrix fibrosis and serum markers of fibrosis in dilated cardiomyopathy, regardless of the definition used for LVRR 10.1007/s00380-016-0930-y
Cytokine 12-month patterns of serum markers of collagen synthesis, transforming growth factor and connective tissue growth factor are similar in new-onset and chronic dilated cardiomyopathy in patients both with and without cardiac fibrosis. pubmed:28460256
Advanced Healthcare Materials Prognostic value of fibrosis-related markers in dilated cardiomyopathy: A link between osteopontin and cardiovascular events. pubmed:29120858
Life Sciences Sustained β-AR stimulation induces synthesis and secretion of growth factors in cardiac myocytes that affect on cardiac fibroblast activation Pubmed:29107793
International Journal of Psychophysiology  Substrate Stiffness Influences the Time Dependence of CTGF Protein Expression in Müller Cells Pubmed:30123889
Journal of Cellular Physiology TGFβ mediates collagen production in human CRSsNP nasal mucosa‐derived fibroblasts through Smad2/3‐dependent pathway and CTGF induction and secretion Pubmed: 30426494
Journal of Clinical Medicine Combination of Plasma Biomarkers and Clinical Data for the Detection of Myocardial Fibrosis or Aggravation of Heart Failure Symptoms in Heart Failure with … Pubmed: 30413105
Nature Communications Macrophage hypoxia signaling regulates cardiac fibrosis via Oncostatin M Pubmed: 31249305
Cardiology Circulating Connective Tissue Growth Factor Is Associated with Diastolic Dysfunction in Patients with Diastolic Heart Failure Pubmed: 31466059
Stem Cells International CTGF Attenuates Tendon-Derived Stem/Progenitor Cell Aging Pubmed: 31827530
Pharmaceuticals Small-Dose Sunitinib Modulates p53, Bcl-2, STAT3, and ERK1/2 Pathways and Protects against Adenine-Induced Nephrotoxicity Pubmed: 33212804
BMC Veterinary Research Lysophosphatidic acid as a regulator of endometrial connective tissue growth factor and prostaglandin secretion during estrous cycle and endometrosis in the … Pubmed: 32943074
Revista Portuguesa de Cardiologia MicroRNAs and ventricular remodeling in aortic stenosis Pubmed: 32682570
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