Mouse Protein Kinase, cGMP Dependent Type I ELISA Kit

From Antibodies.com, this sandwich ELISA kit is designed for the in vitro quantitative determination of mouse Protein Kinase, cGMP Dependent Type I (cGKI) in tissue homogenates or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, using an antibody specific for cGKI that has been pre-coated onto a 96-well microplate. Standards and test samples are added to the wells, allowing cGKI present in each sample to be captured by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-cGKI Antibody is added, which binds the captured cGKI in each well. An HRP-Avidin conjugate is then applied, and TMB substrate solution is used to visualise the HRP enzymatic reaction; the colour shifts to yellow upon addition of acidic stop solution, with the density of yellow being proportional to the amount of cGKI captured. Absorbance is read at 450 nm in a colorimetric microplate reader and the concentration of cGKI is calculated by reference to the standard curve.

Reported sensitivity is <0.059 ng/ml, with an assay range of 0.156-10 ng/ml, and a total assay time of 4h 30m. Sample recovery has been validated in serum (average 91%, range 80-102%), EDTA plasma (average 90%, range 81-100%), and heparin plasma (average 84%, range 80-89%), with n=5 for each matrix; per-dilution recovery is also reported for 1:2, 1:4, 1:8, and 1:16 sample dilutions. 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 5 adhesive plate sealers. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.