CTGF (Full+N-terminal region) ELISA Kit Wako Technical Information and Data

Product Information

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.

CTGF contains four domains, modules 1-4. The N-terminal region (modules 1 and 2) is known as a candidate biomarker of idiopathic pulmonary fibrosis (IPF)¹⁾. However, it has been difficult to accurately measure only the N-terminal region of CTGF, because the N-terminal region and platelet-derived full-length CTGF are both present in the blood.

CTGF (Full + N-terminal region) ELISA Kit Wako is an ELISA kit for detection of full-length CTGF and N-terminal region of CTGF using monoclonal antibodies that recognize CTGF Module 1 and Module 2. The amount of N-terminal CTGF in blood can be measured when this product is used in combination with a kit for detecting full-length CTGF (CTGF (Full) ELISA Kit Wako (Catalog ID 290-84701))²⁾.

Kit Performance

Analyte	CTGF (full-length) and CTGF (N-terminal region)
Sample	Human serum/plasma(EDTA) * heparin plasma is not recommended
Calibration Curve Range	7.81 - 500 pM
Sample Amount Required	Human serum: 5 μL Human plasma (EDTA): 10 μL
Assay Time	2 hours and 50 minutes
Detection Method	Colorimetry

Principle

Data

Spiked recovery test

Sample	Amount spiked (pM)	Measured value (pM)	Amount recovered (pM)	Recovery rate (%)
Human Serum (ID: No.15)	0	61.5	-	-
	10	71.4	9.90	99.0
	50	110	48.5	97.0
	100	163	102	102
	150	215	154	103
	300	370	309	103
	Average			101
Human Plasma (EDTA) (ID: No.15)	0	71.5	-	-
	10	80.8	9.30	93.0
	50	121	49.5	99.0
	100	174	103	103
	150	227	156	104
	300	380	309	103
	Average			100

Dilution linearity test

Human Serum

Human Plasma

Cross-reactivity

Cross-reactivity with each CTGF fragments

Result: 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).

CCN Family	Cross-reactivity (%)
CCN1	N.D.
CCN2 (CTGF)	100
CCN3	N.D.
CCN4	N.D.
CCN5	N.D.
CCN6	N.D.

Result: CTGF (Full + N-terminal region) ELISA Kit Wako has 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.

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 (Catalog ID 290-84701), 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 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: The N-terminal CTGF levels in plasma in IPF patients were significantly higher than those in healthy subjects.