Rat Cardiotrophin Like Cytokine Factor 1 ELISA Kit

From Antibodies.com, this sandwich ELISA kit is intended for the in vitro quantitative determination of rat NNT1 (Cardiotrophin Like Cytokine Factor 1) in serum, plasma or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, using an antibody specific for NNT1 pre-coated onto a 96-well microtiter plate. Standards and test samples are added, and following incubation and washing the wells are incubated with a Biotinylated Anti-NNT1 Antibody. An HRP-Avidin conjugate is then added, and TMB substrate is used to visualise the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of acidic stop solution. The absorbance is read at 450 nm in a microplate reader and the NNT1 concentration calculated by reference to the standard curve.

Key performance specifications include an assay range of 31.2-2000 pg/ml and a sensitivity of <12.9 pg/ml. The colourimetric assay is run on a pre-coated microplate (12 x 8 well strips) with a total assay time of 4h 30m. Kit components comprise a lyophilised 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 plate sealers (5 adhesive strips), with storage at +4°C or -20°C as instructed and the recommendation not to use past the kit's expiration date.

Reported spike-and-recovery values are: serum 80-102% (average 91%, n=5), EDTA plasma 81-100% (average 90%, n=5), and heparin plasma 80-89% (average 84%, n=5). Linearity-of-dilution results at 1:2, 1:4, 1:8 and 1:16 are 87-91%, 87-107%, 74-101% and 92-97% for serum (n=5); 90-105%, 84-101%, 90-101% and 79-108% for EDTA plasma (n=5); and 84-95%, 92-105%, 82-105% and 89-91% for heparin plasma (n=5).