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CTGF(Full) ELISA Kit Wako

for Immunochemistry
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at 2-10 degrees C.
  • Structural Formula
  • Label
  • Packing
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Product Number
Package Size
Price
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Distributor
290-84701
Barcode No
4548995091635
96Tests
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In stock in Japan

Document

Product Specification Sheet
Package Insert
Spectral Data
Certificate of Analysis
Calibration Certificate

Kit component

96 tests

Antibody-coated Plate 1 plate
CTGF Standard 1 vial
Buffer 20 mL
Biotin-conjugated Antibody Solution 100 μL
Peroxidase-conjugated Streptavidin Solution 100 μL
TMB Solution 12 mL
Stop Solution 12 mL
Wash Solution (10×) 100 mL
Standard Sample Buffer 100 mL
Plate Seal 4 sheets

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.

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CTGF (Full) ELISA Kit Wako is an ELISA kit for detection of full-length CTGF using monoclonal antibodies that recognize CTGF Module 1 and Module 4. The amount of N-terminal CTGF in blood can be measured when this product is used in combination with a kit for detection of full-length CTGF and N-terminal region of CTGF (CTGF (Full + N-terminal region) ELISA Kit Wako, Code No. 292-84901)2).

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  • Kit Performance

    Analyte CTGF (full-length)
    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

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Data

Spiked recovery test

Sample Amount spiked
(pM)
Measured value
(pM)
Amount recovered
(pM)
Recovery rate
(%)
Human
Serum
(ID: No.16)
0 37.8 - -
10 47.0 9.20 92.0
50 87.4 49.6 99.2
100 141 103 103
150 191 153 102
300 343 305 102
Average 99.6
Human
Plasma
(EDTA)
(ID: No.16)
0 16.9 - -
10 26.2 9.30 93.0
50 65.6 48.7 97.4
100 122 105 105
150 173 156 104
300 326 309 103
Average 100

Dilution linearity test

  • Human Serum

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  • Human Plasma

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Cross-reactivity

  • Cross-reactivity with each CTGF fragments


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    [Result]
    CTGF (Full) ELISA Kit Wako specifically detected the full-length CTGF.

  • 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 <0.5
    CCN2 (CTGF) 100
    CCN3 <0.5
    CCN4 <0.5
    CCN5 <0.5
    CCN6 <0.5

    [Result]
    CTGF (Full) 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.

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[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 (Code No. 292-84901) 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.

  • 04430763_img08.png
  • 04430763_img07.png
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[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.

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[Result]
The N-terminal CTGF levels in plasma in IPF patients were significantly higher than those in healthy subjects.

References

FAQ

About sample preparation

What anticoagulant should be used for blood samples?
The literature referenced below indicates that the values for N-terminal CTGF are higher when using heparin plasma than when using EDTA or citrate plasma. Heparin should be avoided as an anticoagulant.
[Reference]
Miyazaki, O. et al.: Ann. Clin. Biochem., 47, 205(2010).

About the kit

Are the antibodies used in this kit available for purchase?
Anti CTGF Module 1 monoclonal antibody (30D2) (Product Number: 018-27423)
The detection antibody is not sold separately. The antibody for Module 4 (Product Number: 019-27453) is available, but the clone is different from the one used for the antibody in this kit.

How is the standard quantified?
The standard is prepared and quantified according to the following literature.
[Reference]
Miyazaki, O. et al.: Ann. Clin. Biochem., 47, 205(2010).

About kit usage

What reagents, instruments, and equipment are required for the assay using this kit?
  • Purified water(distilled water)
  • Test tubes for dilution of standard solution and samples
  • Glass utensils for dilution of Wash Solution (graduated cylinder, beaker)
  • Pipettes with disposable tips (one capable of pipetting 10μL of liquid accurately and one capable of pipetting 200 to 500 μL)
  • Repeater pipette, capable of repeatedly dispensing 100μL
  • Water-absorbent material such as a paper towel (to remove any solution remaining on the plate after washing)
  • Vortex-type mixer
  • Microplate shaker (range approx. 500 to 800 rpm)
  • Automatic washer for 96-well plate (if available) or washing bottle
  • Microplate reader capable of measuring at 450 ± 10 nm, with the correction wavelength set at 600 to 650 nm
  • Software for data analysis
Is it necessary to always use this kit with the CTGF (Full + N-terminal region) ELISA Kit Wako (Code No. 292-84901)?
Both this kit and the CTGF (Full + N-terminal region) ELISA Kit Wako are required only when measuring the N-terminal CTGF (Module 1 - 2). If only full length CTGF is measured, this kit alone can be used.
How many samples can be measured using the 96 tests?
Since 16 wells are needed for standards (n = 2), 80 samples (n = 1) or 40 samples (n = 2) can be measured.
Can the kit be divided for use?
Yes, it can. The plate can be cut at every eight-well boundary, and lots are available for standard use. It should be noted that fewer samples can be analyzed when the kit is divided for use, because a calibration curve needs to be drawn each time.

Overview / Applications

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For research use or further manufacturing use only. Not for use in diagnostic procedures.

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