ELISA Kit for Connective Tissue Growth Factor (CTGF) Sus scrofa; Porcine (Pig) Sandwich ELISA

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

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  • ELISA Kit for Connective Tissue Growth Factor (CTGF) Packages (Simulation)
  • ELISA Kit for Connective Tissue Growth Factor (CTGF) Packages (Simulation)
  • ELISA Kit for Connective Tissue Growth Factor (CTGF) Results demonstration
  • SEA010Po.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 88-101 92
EDTA plasma(n=5) 79-98 94
heparin plasma(n=5) 89-103 96

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) 86-93% 91-104% 92-99% 84-96%
EDTA plasma(n=5) 80-90% 82-102% 88-96% 91-98%
heparin plasma(n=5) 89-98% 90-97% 96-105% 78-90%

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.

ELISA Kit for Connective Tissue Growth Factor (CTGF)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Connective Tissue Growth Factor (CTGF). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Connective Tissue Growth Factor (CTGF). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Connective Tissue Growth Factor (CTGF), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Connective Tissue Growth Factor (CTGF) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Promotion of bone formation by naringin in a titanium particle-induced diabetic murine calvarial osteolysis modelPubMed: 19824055
  • Connective tissue growth factor hammerhead ribozyme attenuates human hepatic stellate cell functionPubMed: PMC2726461
  • Activation of JNK Signaling Mediates Connective Tissue Growth Factor Expression and Scar Formation in Corneal Wound HealingPlosone: 0032128
  • Clinical significance of inflammatory and fibrogenic cytokines in diabetic nephropathyScienceDirect: S0009912012000987
  • Connective tissue growth factor (CTGF/CCN2): A protagonist in cardiac allograft vasculopathy development?PubMed: S1053249812010121
  • The Intervention Effect of Rosiglitozone in Ovarian Fibrosis of PCOS RatsBesjournal: Source
  • Concentrations of connective tissue growth factor in patients with nonalcoholic fatty liver disease: Association with liver fibrosisPubMed: 22846210
  • Clinical significance of connective tissue growth factor in hepatitis B virus-induced hepatic fibrosisPubMed: PMC3351780
  • Multitargeted protective effect of Abacopteris penangiana against carrageenan-induced chronic prostatitis in rats.Pubmed: 24211397
  • Cytokine levels as biomarkers of radiation fibrosis in patients treated with breast radiotherapyRo-journal: Source
  • Knock Out of S1P3 Receptor Signaling Attenuates Inflammation and Fibrosis in Bleomycin-Induced Lung Injury Mice ModelPubmed:Pmc4157792
  • Connective tissue growth factor and cardiac diastolic dysfunction: human data from the Taiwan Diastolic Heart Failure Registry and molecular basis by cellular and animal modelsPubmed:24464932
  • Connective Tissue Growth Factor is a Target of Notch Signaling in Cells of the Osteoblastic LineagePubmed:24792956
  • Disruption of collagen homeostasis can reverse established age-related myocardial fibrosisPubmed:25701883
  • Beneficial effects of combined ursodeoxycholic acid and angiotensin-II type 1 receptor blocker on hepatic fibrogenesis in a rat model of nonalcoholic steatohepatitisPubMed: 26190501
  • Suppression of connective tissue growth factor mediates the renoprotective effect of Sitagliptin rather than Pioglitazone in type 2 diabetes mellitusPubmed:27049870
  • TGF-β and Physiological Root Resorption of Deciduous Teeth.pubmed:28035998
  • Fibrosis of extracellular matrix is related to the duration of the disease but is unrelated to the dynamics of collagen metabolism in dilated cardiomyopathypubmed:27516211
  • 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 LVRRpubmed:28004175
  • The relationship between anti-vascular endothelial growth factor and fibrosis in proliferative retinopathy: clinical and laboratory evidencepubmed:27531356
  • Molecular mechanisms underlying fibrosis and elastin destruction in childhood interstitial lung diseasespubmed:27686729
  • Cardioprotection against Heart Failure by Shenfu Injection via TGF-β/Smads Signaling Pathwaypubmed:28698735
  • 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 LVRR10.1007/s00380-016-0930-y
  • 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
  • Prognostic value of fibrosis-related markers in dilated cardiomyopathy: A link between osteopontin and cardiovascular events.pubmed:29120858
  • Sustained β-AR stimulation induces synthesis and secretion of growth factors in cardiac myocytes that affect on cardiac fibroblast activationPubmed:29107793
  • Substrate Stiffness Influences the Time Dependence of CTGF Protein Expression in Müller CellsPubmed:30123889
  • TGFβ mediates collagen production in human CRSsNP nasal mucosa‐derived fibroblasts through Smad2/3‐dependent pathway and CTGF induction and secretionPubmed: 30426494
  • 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
  • Macrophage hypoxia signaling regulates cardiac fibrosis via Oncostatin MPubmed: 31249305
  • Circulating Connective Tissue Growth Factor Is Associated with Diastolic Dysfunction in Patients with Diastolic Heart FailurePubmed: 31466059
  • CTGF Attenuates Tendon-Derived Stem/Progenitor Cell AgingPubmed: 31827530
  • Small-Dose Sunitinib Modulates p53, Bcl-2, STAT3, and ERK1/2 Pathways and Protects against Adenine-Induced NephrotoxicityPubmed: 33212804
  • Lysophosphatidic acid as a regulator of endometrial connective tissue growth factor and prostaglandin secretion during estrous cycle and endometrosis in the …Pubmed: 32943074
  • MicroRNAs and ventricular remodeling in aortic stenosisPubmed: 32682570
  • Manuscript title Microparticles Containing MiR-625-5p In Coronary Artery Reduce Myocardial Fibrosis After Myocardial Infarction
  • High carbohydrate high fat diet causes arterial hypertension and histological changes in the aortic wall in aged rats: The involvement of connective tissue growth …34455071
  • Hepatocyte-specific Smad4 deficiency alleviates liver fibrosis via the p38/p65 pathway

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