CTGF ELISA Kits
Connective Tissue Growth Factor (CTGF) is a secreted protein of approximately 38 kDa produced by umbilical vein and vascular endothelial cells. It is involved in cell adhesion and the proliferation and differentiation of chondrocytes. It is also considered a key factor in tissue fibrosis and reported as a potential marker of fibrosis.
Fujifilm Wako has developed ELISA kits for the measurement of the full-length CTGF and full-length + N-terminal region of CTGF, respectively. By combining the two ELISA methods, the N-terminal region in CTGF in blood can be accurately measured.
About CTGF
Connective Tissue Growth Factor (CTGF) is a secreted protein of approximately 38 kDa produced by umbilical vein and vascular endothelial cells. It is a member of the CCN family (Cyr61/CTGF/Nov), and also called CCN2. There are four major domains in CTGF, Modules 1 to 4. These modules are also called the IGF-binding domain, VWC domain, TSP-1 domain, and CT domain, respectively. Each domain functions by binding different factors.
CTGF plays an important role in tissue fibrosis and has been reported as a potential biomarker for fibrosis. CTGF is induced by TGFβ and contributes to fibrosis by promoting differentiation of myofibroblasts, which produce extracellular matrix, and collagen synthesis1). In mouse skin fibrosis, TGFβ and CTGF are important in the onset and persistence of fibrosis, respectively2). Other reports indicate that CTGF is involved in chondrocyte proliferation, differentiation, and cell adhesion.
In addition to the full-length CTGF, there are also the N-terminal (Modules 1 - 2) and C-terminal (Modules 3 - 4) CTGF fragments, resulting from cleavage between Module 2 and Module 3. The C-terminal CTGF is susceptible to degradation, and CTGF forms present in the blood are thought to be primarily the full-length and the N-terminal CTGF.
Although the N-terminal CTGF is a potential biomarker for idiopathic pulmonary fibrosis (IPF)3), it has been difficult to measure only the N-terminal CTGF when the full-length and N-terminal CTGF are both present.
CTGF ELISA Kit Wako
Fujifilm Wako has developed ELISA kits for the measurement of the full-length CTGF and full-length + N-terminal region of CTGF, respectively. By combining the two ELISA methods, the N-terminal region in CTGF in blood, expected to be potential biomarker for idiopathic pulmonary fibrosis (IPF), can be accurately measured4).
Product Lineup
| Product Name | CTGF(Full) ELISA Kit Wako | CTGF(Full + N-terminal region) ELISA Kit Wako |
|---|---|---|
| Product Number | 290-84701 | 292-84901 |
| Analyte | CTGF (full-length) | CTGF (full-length) and CTGF (N-terminal region) |
| Principle |
 |
 |
| Analysis Sampe | Human serum, Human plasma (EDTA) | |
| Calibration Curve Range |
7.81 - 500 pM | |
| Sample Volume | Human serum: 5 μL Human plasma (EDTA): 10 μL |
|
| Measurement Duration | 2 hours and 50 minutes | |
| Detection Method | Colorimetry | |
Performance Data
Cross-reactivity with each CTGF fragments
Cross-reactivity with the full-length CTGF and N- and C-terminal fragments was measured.
[Result]
CTGF (Full) ELISA Kit Wako specifically detected the full-length and CTGF (Full + N-terminal region) ELISA Kit Wako specifically detected the full-length and N-terminus.
Cross-reactivity with CCN family proteins
Cross-reactivity with CCN family proteins, including CTGF, was determined (500 pM, n=8).
| CTGF ELISA (Full) Cross-reactivity (%) |
CTGF ELISA (Full + N-terminal region) Cross-reactivity (%) |
|
|---|---|---|
| CCN1 | <0.5 | N.D. |
| CCN2 (CTGF) |
100 | 100 |
| CCN3 | <0.5 | N.D. |
| CCN4 | <0.5 | N.D. |
| CCN5 | <0.5 | N.D. |
| CCN6 | <0.5 | N.D. |
[Result]
FUJIFILM Wako’s kits had high specificity for CCN2 (CTGF) among the CCN family proteins.
Comparison of CTGF reactivity between each products
Recombinant human CTGF protein (rHuman CTGF) with a known concentration (1) Full-length, (2) N-terminal region, and (3) C-terminal region were serially diluted and measured with each products. The measurement procedure followed the instruction manual of each products.
[Result]
CTGF (full length + N-terminal region) ELISA Kit Wako showed high reactivity to CTGF (full length) and CTGF (N-terminal region), and CTGF (full length) ELISA Kit Wako to CTGF (full length). The kit (competitor R) showed high reactivity only with CTGF (full length). The kit (competitor M, N-terminal region specific) hardly reacted with any of the CTGF in our examination.
Application Data
Measurement of the N-terminal region (dilution linearity test)
Full-length plus N-terminal region of CTGF and the only full-length CTGF were measured using the CTGF (Full + N-terminal region) ELISA Kit Wako and the CTGF (Full) ELISA Kit Wako, respectively. The amount of CTGF (N-terminal region) was calculated from the difference between the two measurements, and dilution linearity was determined.
[Result]
The amount of CTGF (N-terminal region) measured using this method showed excellent dilution linearity.
Measurement of N-terminal CTGF in plasma from healthy subjects and IPF patients
The Full-length CTGF and N-terminal CTGF levels were determined in EDTA plasma collected from healthy subjects and patients with idiopathic pulmonary fibrosis (IPF) using the CTGF (Full + N-terminal region) ELISA Kit Wako and CTGF (Full) ELISA Kit Wako.
[Result]
In plasma, the full-length CTGF showed no significant difference between healthy subjects and IPF patients. On the other hand, the N-terminal CTGF levels of IPF patients was significantly higher than that of healthy subjects.
References
- Biernacka, A., Dobaczewski, M., and Frangogiannis, N. G.: Growth Factors, 29(5), 196(2011).
TGF-β signaling in fibrosis - Mori T., et al.: J. Cell Physiol., 181(1), 153(1999).
Role and interaction of connective tissue growth factor with transforming growth factor-beta in persistent fibrosis: A mouse fibrosis model. - Kono, M. et al.: Clin. Chim. Acta, 412, 2211(2011).
Plasma CCN2 (connective tissue growth factor; CTGF) is a potential biomarker in idiopathic pulmonary fibrosis (IPF) - Miyazaki, O. et al.: Ann. Clin. Biochem., 47, 205(2010).
Subtraction method for determination of N-terminal connective tissue growth factor
Product List
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CTGF (Full) ELISA Kit
CTGF (Full + N-terminal region) ELISA Kit
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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