CTGF ELISA Kit

Catalog No. ABIN6730903

Product Details CTGF ELISA Kit

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

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 7.8 pg/mL - 500 pg/mL

Minimum Detection Limit: 7.8 pg/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of CTGF in human serum, plasma, tissue homogenates.

We offer validation data (WB) for the kit components. So you can be sure to order a reliable ELISA kit product composed of high quality reagents.

Sample Type: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Connective Tissue Growth Factor (CTGF)

Cross-Reactivity (Details): No significant cross-reactivity or interference between Connective Tissue Growth Factor (CTGF) and analogues was observed.

Sensitivity: 3.5 pg/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol:

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.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 5,000pg/mL. Firstly dilute the stock solution to 500pg/mL and the diluted standard serves as the highest standard (500pg/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/mL, 7.8pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Storage: 4 °C/-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Expiry Date: 6 months

References for CTGF ELISA Kit (ABIN6730903)

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Rubiś, Wiśniowska-Smiałek, Wypasek, Rudnicka-Sosin, Hlawaty, Leśniak-Sobelga, Kostkiewicz, Podolec et al.: "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 ..." in: Cytokine, Vol. 96, pp. 217-227, (2018) (PubMed).

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Hamed, El-Saied, Saad, Yousef, Mohamed, Sabry: "Molecular mechanisms underlying fibrosis and elastin destruction in childhood interstitial lung diseases." in: Pathophysiology : the official journal of the International Society for Pathophysiology, Vol. 23, Issue 4, pp. 275-283, (2016) (PubMed).

Rubiś, Wiśniowska-Śmialek, Wypasek, Biernacka-Fijalkowska, Rudnicka-Sosin, Dziewiecka, Faltyn, Khachatryan, Karabinowska, Kozanecki, Tomkiewicz-Pająk, Podolec: "Fibrosis of extracellular matrix is related to the duration of the disease but is unrelated to the dynamics of collagen metabolism in dilated cardiomyopathy." in: Inflammation research : official journal of the European Histamine Research Society ... [et al.], Vol. 65, Issue 12, pp. 941-949, (2016) (PubMed).

Rubiś, Wiśniowska-Śmiałek, Biernacka-Fijałkowska, Rudnicka-Sosin, Wypasek, Kozanecki, Dziewięcka, Faltyn, Karabinowska, Khachatryan, Hlawaty, Leśniak-Sobelga, Kostkiewicz, Płazak, Podolec: "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 LVRR." in: Heart and vessels, Vol. 32, Issue 6, pp. 714-725, (2016) (PubMed).

Westbury, Haviland, Davies, Gothard, Abdi, Sydenham, Bowen, Stratton, Short, Yarnold: "Cytokine levels as biomarkers of radiation fibrosis in patients treated with breast radiotherapy." in: Radiation oncology (London, England), Vol. 9, pp. 103, (2015) (PubMed).

Rosin, Sopel, Falkenham, Lee, Légaré: "Disruption of collagen homeostasis can reverse established age-related myocardial fibrosis." in: The American journal of pathology, Vol. 185, Issue 3, pp. 631-42, (2015) (PubMed).

Wu, Wang, Lee, Chang, Su, Yeh, Su, Chen, Chiang, Hwang, Lin, Tsai: "Connective tissue growth factor and cardiac diastolic dysfunction: human data from the Taiwan diastolic heart failure registry and molecular basis by cellular and animal models." in: European journal of heart failure, Vol. 16, Issue 2, pp. 163-72, (2014) (PubMed).

Shi, Chang, Yang, Zhang, Yu, Wu: "Activation of JNK signaling mediates connective tissue growth factor expression and scar formation in corneal wound healing." in: PLoS ONE, Vol. 7, Issue 2, pp. e32128, (2012) (PubMed).

El Mesallamy, Ahmed, Bassyouni, Ahmed: "Clinical significance of inflammatory and fibrogenic cytokines in diabetic nephropathy." in: Clinical biochemistry, Vol. 45, Issue 9, pp. 646-50, (2012) (PubMed).

Colak, Senates, Coskunpinar, Oltulu, Zemheri, Ozturk, Doganay, Mesci, Yilmaz, Enc, Kiziltas, Ulasoglu, Tuncer: "Concentrations of connective tissue growth factor in patients with nonalcoholic fatty liver disease: association with liver fibrosis." in: Disease markers, Vol. 33, Issue 2, pp. 77-83, (2012) (PubMed).

Piao, Brigstock, Zhu, Zhang, Gao: "Clinical significance of connective tissue growth factor in hepatitis B virus-induced hepatic fibrosis." in: World journal of gastroenterology : WJG, Vol. 18, Issue 18, pp. 2280-6, (2012) (PubMed).

Pantou, Manginas, Alivizatos, Degiannis: "Connective tissue growth factor (CTGF/CCN2): a protagonist in cardiac allograft vasculopathy development?" in: The Journal of heart and lung transplantation : the official publication of the International Society for Heart Transplantation, Vol. 31, Issue 8, pp. 881-7, (2012) (PubMed).

Zhou, Zhang, Zhang, Zhu: "Promotion of bone formation by naringin in a titanium particle-induced diabetic murine calvarial osteolysis model." in: Journal of orthopaedic research : official publication of the Orthopaedic Research Society, Vol. 28, Issue 4, pp. 451-6, (2010) (PubMed).

Gao, Brigstock: "Connective tissue growth factor hammerhead ribozyme attenuates human hepatic stellate cell function." in: World journal of gastroenterology : WJG, Vol. 15, Issue 30, pp. 3807-13, (2009) (PubMed).

Maruyama, Fujita, Taniguchi: "Sequence of a cDNA coding for human IRF-1." in: Nucleic acids research, Vol. 17, Issue 8, pp. 3292, (1989) (PubMed).

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Target Details for CTGF

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

Abstract: 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

UniProt: P29279

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