Mouse Sestrin 3 ELISA Kit

Provided by Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is intended for the in vitro quantitative determination of mouse SESN3 (Sestrin 3) in tissue homogenates or other biological fluids. An antibody specific for SESN3 is pre-coated onto a 96-well microplate formatted as 12 x 8 well strips. Standards and samples are added; SESN3 is bound by the immobilised antibody and detected sequentially with a biotinylated anti-SESN3 antibody, an HRP-Avidin conjugate and TMB substrate. The enzymatic reaction yields a blue product that turns yellow after addition of acidic stop solution, and the concentration of SESN3 is calculated from the absorbance read at 450 nm against a standard curve.

The assay has a working range of 0.156-10 ng/ml and a sensitivity of <0.051 ng/ml, with a typical run time of 4 hours 30 minutes. Validation across serum (n=5), EDTA plasma (n=5) and heparin plasma (n=5) reports mean recoveries of 91%, 90% and 84% respectively, with individual recovery ranges of 80-102% (serum), 81-100% (EDTA plasma) and 80-89% (heparin plasma). Linearity at dilutions of 1:2, 1:4, 1:8 and 1:16 is also reported for each matrix.

Components supplied include a pre-coated 96-well microplate, lyophilised standard, detection solutions A and B, sample dilution buffer, wash buffer (30X), TMB substrate, stop solution and adhesive plate sealers. Storage temperatures vary by component: the microplate, standards and detection solutions are stored at -20°C, while wash buffer, TMB substrate and stop solution are stored at +4°C. The kit should not be used past its expiration date.