| Fibronectin (FN) also known as LETS, is identified on the surface FN of fibroblasts by labeling with radioactive compounds or specific antibodies. Fibronectin is a 430kD dimeric glycoprotein that exists in 2 forms, termed cellular and plasma fibronectin. Cellular and plasma fibronectins are heterodimers consisting of similar but not identical polypeptides. These two forms of FN differ in biologic activity. Fibronectins bind cell surface FNs and various compounds including collagen, fibrin, heparin, DNA, and actin. Because fibronectin stimulates endocytosis in several systems and promotes the clearance of particulate material from the circulation, it could function in the clearance of C1q-coated material such as immune complexes or cellular debris. Fibronectins are involved in cell adhesion, cell motility, opsonization, would healing, and maintenance of cell shape. LETS, encoded on chromosome 8, is responsible for the LETS protein expression in humans. Because LETS has been implicated in tumorigenicity and cellular transformation, it is of interest that rearrangement or modifications in the number of chromosome 8 have been associated with certain forms of cancer. The standard used in this kit is isolated from human plasma with the molecular mass of 200-250kD. Intended Use: Sandwich ELISA quantitative detection kit for human Fibronectin in cell culture supernates, serum and plasma (heparin, EDTA, citrate). Range: 156-10,000pg/ml Sensitivity: <10pg/ml Specificity: Natural human Fibronectin Crossreactivity: No detectable crossreactivity with other relevant proteins Kit Components: 143835A: Microtiter Strip, 1x96 wells 143835B: Fibronectin Standard, 2x10ng 143835C: Anti-human Fibronectin (Biotin), 1x130ul 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml Storage and Stability: Store *143835B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add 100ul of samples and standards and incubate the plate at 37C for 90 min. Do not wash. 2. Add 100ul biotinylated antibody and incubate the plate at 37C for 60 min. Wash plate 3 times with 0.01M PBS or TBS. 3. Add ABC working solution and incubate the plate at 37C for 30 minutes. Wash plate 5 times with 0.01M PBS or TBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 25-30 minutes. 5. Add 100ul TMB stop solution and read. |
