Rat Visfatin ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com designed for the in vitro quantitative determination of rat Visfatin in serum, plasma, tissue homogenates or other biological fluids. The kit employs a 96-well microtiter plate pre-coated with an antibody specific for Visfatin. Standards and test samples are added into the wells and Visfatin present in each sample is bound by the immobilized antibody. Following incubation, the wells are washed and incubated with a Biotinylated Anti-Visfatin Antibody, which binds the captured Visfatin. An HRP-Avidin conjugate is then added and the plate is incubated. TMB substrate solution is used to visualize the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow after addition of acidic stop solution. The intensity of yellow is proportional to the amount of Visfatin captured in each well. The concentration of Visfatin is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

The assay has a stated sensitivity of 0.55 ng/ml and a product range of 1.56-100 ng/ml. Assay time is 4h 30m, with detection on a Colorimetric Microplate Reader. Reported recovery values are: Serum (n=5) range 80%-102%, average 91%; EDTA Plasma (n=5) range 81%-99%, average 90%; Heparin Plasma (n=5) range 80%-89%, average 84%. Linearity at dilutions of 1:2, 1:4, 1:8 and 1:16 is reported for Serum, EDTA Plasma and Heparin Plasma. Kit components include a Pre-Coated 96 Well Microplate (12 x 8 Well Strips), Lyophilized Standard (2 vials), Detection Solution A (120 µl), Detection Solution B (120 µl), Wash Buffer 30X (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml) and Plate Sealers (5 adhesive strips). Components should be stored at +4°C or -20°C as instructed.