Human Signal Transducer And Activator Of Transcription 1 ELISA Kit

Designed for the in vitro quantitative measurement of human Signal Transducer and Activator of Transcription 1 (STAT1) in tissue homogenates, cell lysates and other biological fluids, this Antibodies.com sandwich enzyme-linked immunosorbent assay (sELISA) kit uses a pre-coated microtiter plate to which an anti-STAT1 capture antibody has been immobilised [S1]. Standards and test samples are added to the wells and STAT1 present in each sample binds to the immobilised antibody [S1]. After incubation and washing, a biotinylated anti-STAT1 detection antibody is added, followed by an HRP-avidin conjugate [S1]. TMB substrate is then used to visualise the HRP enzymatic reaction, producing a blue product that turns yellow after addition of acidic stop solution [S1]. The density of yellow is proportional to the amount of STAT1 captured, and the concentration is calculated by reading absorbance at 450 nm in a colorimetric microplate reader against a standard curve [S1].

Reported sensitivity is less than 0.114 ng/ml with a measuring range of 0.312–20 ng/ml, and a representative run takes 4 hours 30 minutes [S1]. Spike-and-recovery testing gave averages of 91% in serum (range 80%–102%, n=5), 90% in EDTA plasma (range 81%–100%, n=5) and 84% in heparin plasma (range 80%–89%, n=5) [S1]. Dilution linearity assessed at 1:2, 1:4, 1:8 and 1:16 yielded 87–91%, 87–107%, 74–101% and 92–97% in serum (n=5); 90–105%, 84–101%, 90–101% and 79–108% in EDTA plasma (n=5); and 84–95%, 92–105%, 82–105% and 89–91% in heparin plasma (n=5) [S1]. Components are stored at +4°C or –20°C as instructed and the kit should not be used past its expiration date [S1].