CD147, a member of the immunoglobulin superfamily, is a transmembrane glycoprotein included two immunoglobulin-like domains 1). CD147 is also known as extracellular matrix metalloproteinase inducer (EMMPRIN) or basigin and is enriched on the surface of many malignant tumor cells and stromal cells. The up-regulation of the matrix metalloproteinases (MMPs) by CD147 is believed to be progression of malignancies through tumor growth, invasion and metastasis 2,3,4). In the recent studies, EMMPRIN fragment (22KDa) is released in a soluble form by the protelytic cleavage of MMPs from tumor cell surface. There are several evidences that the soluble CD147 is existed into the culture medium of several tumor cells 5,6).
This assay kit employs the quantitative sandwich ELISA technique based on two mouse monoclonal antibodies against to specific different epitope of EMMPRIN. One of specific antibody is pre-coated onto 96-well microplate.
Calibrators and samples (culture medium and clinical sample, e.g. urine) are applied into the wells and incubated, EMMPRIN is captured by the coated antibody. Following, the microplate is incubated with biotinylated antibody. And next step, HRP-conjugated streptavidin is added to the well and incubated. Finally, a chromogenic substrate solution (H2O2 and OPD) is applied to each test well and color develops in proportion to EMMPRIN level. The enzymatic reaction is stopped by addition of sulphuric acid and absorbance of the color intensity is measured spectrophotometrically at 490 nm. Calibration curve is constructed, and EMMPRIN concentrations of samples are able to determine using the calibration curve.
< KIT PERFORMANCE>
Sensitivity (the corresponding concentration of blank absorbance + 2SD); 0.30 ng/mL Intra assay Precision (n = 10); CV < 10% Inter assay Precision (n = 10); CV < 10%
1. Antibody pre-coated 96-well microplate 1 plate (6 strips)
2. Biotinylated antibody, concentrated (100 x) 1 vial (120 L)
3. HRP-conjugated streptavidin, concentrated (100 x) 1 vial (120 L)
4. Antibody diluent 1 vail (30 mL)
5. Wash solution, concentrated (20 x) 1 vial (30 mL).
6. Substrate solution 1 vial (30 mL)
7. OPD tablet 2 tablets
8. Stop solution (sulphuric acid) 1 vial (15 mL)
9. Sample diluent 1 vail (15 mL)
10. EMMPRIN standard (1000 ng/mL) 2 vial (30 L/vial)
Soluble CD147(EMMPRIN) ELISA Kit
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.