Rat Mothers Against Decapentaplegic Homolog 3 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com designed for the in vitro quantitative determination of rat Smad3 (Mothers Against Decapentaplegic Homolog 3) in tissue homogenates, cell lysates and other biological fluids. The product is intended for research use only and is not for diagnostic or therapeutic applications.

The assay uses a pre-coated microplate configured as 12 x 8 well strips and the sandwich immunoassay technique, in which captured Smad3 is bound by a biotinylated anti-Smad3 detection antibody followed by an HRP-avidin conjugate. Signal is generated with TMB substrate, producing a blue colour that converts to yellow upon addition of the acidic stop solution; absorbance is read at 450 nm in a colorimetric microplate reader. Reported sensitivity is <0.127 ng/ml and the assay range is 0.312–20 ng/ml. Total assay time is stated as 4 h 30 m, and kit components are stored at +4 °C or -20 °C as instructed.

Spike-and-recovery testing reported average recoveries of 91% for serum (range 84–99%, n=5), 92% for EDTA plasma (range 82–103%, n=5) and 93% for heparin plasma (range 84–102%, n=5). Linearity was assessed across 1:2, 1:4, 1:8 and 1:16 dilutions for serum, EDTA plasma and heparin plasma. Required materials not supplied with the kit include a microplate reader capable of measuring absorbance at 450 nm ± 10 nm, multichannel and single-channel pipettes, a 37 °C incubator, and deionised or distilled water.