CTGF ELISA Kit

Catalog No. ABIN956182

Product Details CTGF ELISA Kit

Target: CTGF (Connective Tissue Growth Factor (CTGF))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: This CTGF ELISA kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.

Analytical Method: Quantitative

Characteristics: This CTGF enzyme linked immunosorbent assay applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a polyclonal antibody specific for CTGF. Calibrators or samples are then added to the microtiter plate wells and CTGF if present, will bind to the antibody pre-coated wells. In order to quantitatively determine the amount of CTGF present in the sample, a standardized preparation of horseradish peroxidase (HRP)-conjugated polyclonal antibody, specific for CTGF are added to each well to sandwich the CTGF immobilized on the plate. The microtiter plate undergoes incubation, and then the wells are thoroughly washed to remove all unbound components. Next, A and B substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain CTGF and enzyme-conjugated antibody will exhibit a change in color. The enzyme-substrate reaction is terminated by addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm.

Components: Microtiter Plate: 96 wells
Calibrator 1 (0 pg/mL)
Calibrator 2 (250 pg/mL)
Calibrator 3 (500 pg/mL)
Calibrator 4 (1,000 pg/mL)
Calibrator 5 (2,500 pg/mL)
Calibrator 6 (5,000 pg/mL)
Enzyme Conjugate (1 x 6 mL)
Substrate A (1 x 6 mL)
Substrate B (1 x 6 mL)
Stop Solution (1 x 6 mL)
Wash Buffer (100X concentrate) (1 x 6 mL)
Lysis Buffer Solution (1 x 6 mL)
Note: The lysis buffer solution is used only when the sample is cell culture fluid & body fluid & tissue homogenate, If the sample is serum or blood plasma, then the lysis buffer solution is a superfluous reagent.

Material not included: 1. Microplate reader capable of measuring absorbance at 450 nm.
2. Pipettes and pipette tips.
3. 100 mL and 1 liter graduated cylinders.
4. Calibrated adjustable precision pipettes, preferably with disposable plastic tips. (A manifold multi- channel pipette is desirable for large assays.)
5. 37°C incubator.
6. Absorbent paper.
7. Distilled or de-ionized water
8. Data analysis and graphing software. Graph paper: linear (Cartesian), log-log or semi-log, or log-logit as desired.
9. Tubes to prepare calibrator or sample dilutions.

Application Details

Plate: Pre-coated

Assay Procedure:

It is recommended that all Calibrators and Samples be added in duplicate to the Microtiter Plate.
1. Secure the desired number of coated wells in the holder then add 50 µL of Calibrators or Samples to the appropriate well in the antibody pre-coated Microtiter Plate.
2. Add 100 µL of Conjugate to each well. Mix well. Mixing well in this step is important. Cover and incubate for 1 hour at 37°C.
3. Wash the microtiter plate using one of the specified methods indicated below:
4. Manual Washing: Remove incubation mixture by aspirating contents of the plate into a sink or proper waste container. Fill in each well completely with diluted wash solution, and then aspirate contents of the plate into a sink or proper waste container. Repeat this procedure five times for a total of five washes. After washing, invert plate, and blot dry by hitting the plate onto absorbent paper or paper towels until no moisture appears. Note: Hold the sides of the plate frame firmly when washing the plate to assure that all strips remain securely in frame. Complete removal of liquid at each step is essential to good performance.
5. Automated Washing: Wash plate five times with diluted wash solution (350-400 µL/well/wash) using an auto washer. After washing, dry the plate as above. It is recommended that the washer be set for a soaking time of 10 seconds and shaking time of 5 seconds between each wash.
6. Add 50 µL substrate A and 50 µL substrate B to each well, subsequently. Cover and incubate for 15 minutes at 20-25°C. (Avoid sunlight.)
7. Add 50 µL of stop solution to each well. Mix well.
8. Read the optical density (OD) at 450 nm using a microtiter plate reader immediately.

Calculation of Results:

1. This calibration curve is used to determine the amount of an unknown sample. Construct a calibration curve by plotting the average O.D. (450 nm) for each calibrator on the vertical (Y) axis against the concentration on the horizontal (X) axis, and draw a best fit curve through the points on the graph.
   2. First, calculate the mean O.D. value for each calibrator and sample. All O.D. values are subtracted by the mean value of the blank control before result interpretation. Construct the calibration curve using graph paper or statistical software.
   3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the calibration curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration.
   4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own calibration curve.
   5. The sensitivity in this assay is 1.0 pg/mL.
   6. This assay has high sensitivity and excellent specificity for detection of CTGF. No significant cross- reactivity or interference between CTGF and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between CTGF and all the analogues, therefore, cross reaction may still exist in some cases.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: All reagents provided are stored at 4°C. Refer to the expiration date on the label.

Expiry Date: The expiry date is stated on the label.

Target Details for CTGF

Target: CTGF (Connective Tissue Growth Factor (CTGF))

Alternative Name: CTGF (CTGF Products)

Synonyms: Ccn2 ELISA Kit, Fisp12 ELISA Kit, Hcs24 ELISA Kit, fisp-12 ELISA Kit, CCN2 ELISA Kit, HCS24 ELISA Kit, IGFBP8 ELISA Kit, NOV2 ELISA Kit, CTGRP ELISA Kit, ctgf ELISA Kit, hm:zeh1559 ELISA Kit, si:dkey-266k12.4 ELISA Kit, wu:fl25c03 ELISA Kit, zeh1559 ELISA Kit, zgc:136644 ELISA Kit, XCTGF ELISA Kit, ccn2 ELISA Kit, nov2 ELISA Kit, hcs24 ELISA Kit, igfbp8 ELISA Kit, CTGF ELISA Kit, si:ch211-173b8.2 ELISA Kit, connective tissue growth factor ELISA Kit, connective tissue growth factor a ELISA Kit, connective tissue growth factor L homeolog ELISA Kit, connective tissue growth factor b ELISA Kit, Ctgf ELISA Kit, CTGF ELISA Kit, ctgfa ELISA Kit, ctgf.L ELISA Kit, ctgf ELISA Kit, ctgfb ELISA Kit

Pathways: Regulation of Lipid Metabolism by PPARalpha, Positive Regulation of Endopeptidase Activity, Growth Factor Binding