Human Signal Transducer And Activator Of Transcription 3 ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of human STAT3 in tissue homogenates, cell lysates, cell culture supernates, and other biological fluids. The kit employs the sandwich enzyme immunoassay technique using a 96-well microtiter plate pre-coated with an antibody specific for STAT3. Standards and test samples are added to the wells, where STAT3 is captured by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-STAT3 Antibody is added, followed by HRP-Avidin conjugate and TMB substrate solution. The resulting enzymatic reaction produces a blue-coloured product that turns yellow upon addition of acidic stop solution. The intensity of yellow colour is proportional to the amount of STAT3 captured in each well, and concentration is calculated by reading the O.D. absorbance at 450 nm against a standard curve.

The quantitative sandwich assay has a reported product range of 0.312–20 ng/ml and a sensitivity of <0.117 ng/ml. Detection is colorimetric using a microplate reader. The pre-coated microplate is configured as 12 x 8 well strips, and the total assay time is approximately 4 hours 30 minutes. The kit includes a pre-coated 96-well microplate, lyophilized standard, Detection Solution A, Detection Solution B, Wash Buffer (30X), Sample Dilution Buffer, TMB Substrate, Stop Solution, and plate sealers. Recovery performance has been characterised for serum (average 91%, range 80–102%, n=5), EDTA plasma (average 90%, range 81–100%, n=5), and heparin plasma (average 84%, range 80–89%, n=5). Linearity of dilution has also been demonstrated across serum, EDTA plasma, and heparin plasma at 1:2, 1:4, 1:8, and 1:16 dilutions. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.