RayBio® Phospho-SMAD2 (Ser245/250/255) and Total SMAD2 ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human, mouse and rat cell lysates. By determining phosphorylated SMAD2 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human, mouse and rat phospho-SMAD2 and total SMAD2. An anti-pan SMAD2 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and SMAD2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-SMAD2 (Ser245/250/255) antibody is used to detect phosphorylated SMAD2 or mouse anti-SMAD2 antibody is used to detect pan SMAD2. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-conjugated anti-mouse IgG is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of SMAD2 (Ser245/250/255) or pan SMAD2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4°C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge, and store at –20°C. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C.