| Human Clara Cell Protein ELISA is a sandwich enzyme immunoassay for the quantitative measurement of human Clara Cell Protein in serum and plasma (EDTA, citrate, heparin). |
|
| Catalog # | C5828-02 |
| Background | Human Clara Cell Protein (CC16, CC10 and also called uteroglobin, urinary protein 1 or Clara Cell Secretory Protein) belongs to the family of secretoglobins. It is a secreted protein product of non-ciliated bronchiolar Clara cells. Its function remains to be elucidated. There is convincing data suggesting its phospholipase A2 inhibitory activity as well as a number of other immunomodulatory features including inhibition of interferon gamma signaling and Th1 vs. Th2 lymphocyte regulation. It was proposed as a potential peripheral marker of respiratory epithelial injury and bronchial dysfunction. Clara Cell Protein 16 concentrations have been determined in both serum and bronchoalveolar lavage fluid in numerous studies since 1994. In serum, its increase is associated with age, asbestos, nitrogen chloride and ozone exposure, sarcoidosis and high PEEP ventilation. Decreased serum CC16 levels are found after pulmonary resection, in silica-exposed workers, smokers and in asthma. Decreased CC16 concentrations were also found in the amniotic fluid of fetuses suffering from pulmonary hypoplasia caused by various mechanisms (diaphragmatic hernia, diabetic fetopathy, Turner and Down syndrome). In pleural effusions, the CC16 concentration appears to be associated with its diffusion from the lung as evidenced by high CC16 levels in cardiac pleural congestion. |
| Features | • The total assay time is less than four hours. |
| • The kit measures homodimeric Clara Cell Protein. |
| • Quality Controls are human serum based. No animal sera are used. |
| • Suitable for use with human serum or plasma samples |
| Test Principle | In the Human Clara Cell Protein ELISA, calibrators or samples are incubated with a polyclonal rabbit anti-human Clara Cell Protein antibody coated in microtiter wells. After one-hour incubation and a washing, polyclonal anti-human Clara Cell Protein antibody labeled with biotin is added and incubated with captured Clara Cell Protein. After a thorough wash, streptavidin labeled with HRP is added. After one hour incubation and the last washing step, the remaining conjugate is allowed to react with the substrate H2O2- tetramethylbenzidine. The reaction is stopped by addition of acidic solution. Absorbance of the resulting yellow product is measured at 450nm. The absorbance is proportional to the concentration of Clara Cell Protein. A standard curve is constructed by plotting absorbance values versus Clara Cell Protein concentrations of calibrators. Concentrations of unknown samples are determined using this standard curve. |
| Performance Characteristics | Sensitivity: ~0.04ng/ml |
| The limit of detection is defined as such a concentration of Clara Cell Protein giving absorbance higher than mean absorbance of blank* plus three standard deviations of the absorbance of blank: Ablank + 3*SDblank is better than 0.04ng/ml of sample. |
| Specificity | The antibodies in the Human Clara Cell Protein ELISA kit are highly specific of human Clara Cell Protein with no detectable crossreactivities to the cytokines that may be present in human |
| serum. |
| Range | 2-100ng/ml Clara Cell Protein/ml of sample |
| Sensitivity | ~0.2ng/ml |
| Sample Volume | 100ul/well |
| Assay Time | Total assay time is 3-4 hours. |
| Definition of Clara Cell Protein Master Calibrator | The Clara Cell Protein is purified from human urine and is used as the calibrator. The Clara Cell Protein is a 16kD dimeric protein consisting of two disulfide-linked polypeptide chains. The CC16 concentration strongly depends on the method, which is used for the protein determination. Master standard contains 50ng of CC16 measured by Bradford method (used in this kit), 215ng of CC16 measured by BCA method and 340ng of CC16 measured by Lowry method. |
| Kit Components | C5828-02A: Microtiter Strips, coated with capture polyclonal antibody, sealed, 1x96 wells |
| C5828-02B*: Clara Cell Protein Master Calibrator, 1x1vial |
| C5828-02C*: Quality Controls: High, 15.2-22.8ng/ml, 2x1vial |
| C5828-02D*: Quality Controls: Low, 5.7-8.5ng/ml, 2x1vial |
| C5828-02E: Pab (Biotin), Ready to use, 1x13ml |
| C5828-02F: Streptavidin (HRP), Ready to use, 1x13ml |
| C5828-02G: Dilution Buffer, Ready to use, 1x20ml |
| C5828-02H: Wash Solution, 10X, 1x100ml |
| C5828-02J: TMB Substrate Solution, Ready to use, 1x13ml |
| C5828-02K: Stop Solution, Ready to use, 1x13ml |
| Storage and Stability | Store at 4°C. Stable for 6 months at 4°C. *Note: Store C5828-02B, C5828-02C and C5828-02D at 4°C as a lyophilized powder. Upon reconstitution, aliquot and store the undiluted standard and controls at -20°C. Stable for 3 months. |
| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
