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
Storage and Stability:Store *143835B powder at 4°C. Once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. 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 37°C for 90 min. Do not wash
2. Add 100ul biotinylated antibody and incubate the plate at 37°C for 60 min. Wash plate 3 times with 0.01M PBS or TBS
3. Add ABC working solution and incubate the plate at 37°C for 30 minutes. Wash plate 5 times with 0.01M PBS or TBS
4. Add TMB color developing agent and incubate the plate at 37°C in dark for 25-30 minutes
