Mouse Fibronectin ELISA Kit

Sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com for the in vitro quantitative determination of mouse Fibronectin in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.

The assay uses the sandwich enzyme immunoassay technique. An antibody specific for Fibronectin is pre-coated onto a microplate; standards and test samples are added and Fibronectin present binds to the immobilised antibody. After washing, a Biotinylated Anti-Fibronectin Antibody is added, followed by HRP-Avidin conjugate. TMB substrate solution then visualises the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow upon addition of acidic stop solution. The intensity of the yellow colour is proportional to the amount of Fibronectin captured in the well. Concentration is calculated by reading the O.D. absorbance at 450 nm on a colorimetric microplate reader and referring to the standard curve.

The standard assay range is 0.156–10 ng/ml and sensitivity is <0.062 ng/ml. Assay time is approximately 4 hours 30 minutes, with colorimetric detection. Validation recovery in serum ranged 80–102% (average 91%, n=5); EDTA plasma 81–99% (average 90%, n=5); heparin plasma 80–89% (average 84%, n=5). Dilution linearity was assessed at 1:2, 1:4, 1:8 and 1:16 dilutions in serum, EDTA plasma and heparin plasma (n=5 each). Kit components should be stored at +4°C or –20°C as instructed and the product must not be used past its expiration date.