Rat 5-Hydroxytryptamine Receptor 2C ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (ELISA) is designed for the in vitro quantitative determination of rat 5-Hydroxytryptamine Receptor 2C (5HT2C Receptor) in tissue homogenates or other biological fluids. The assay employs a 96-well microtiter plate pre-coated with an antibody specific for the 5HT2C Receptor. Standards and test samples are added to the wells and bound by the immobilized antibody, followed by incubation with a Biotinylated Anti-5HT2C Receptor Antibody and an HRP-Avidin conjugate. Detection is performed using TMB substrate solution, with absorbance read at 450 nm in a colorimetric microplate reader; the resulting colour density is proportional to the amount of 5HT2C Receptor present in each well.

The reported assay range is 0.312–20 ng/ml, with a sensitivity of 0.083 ng/ml and an assay time of 4 hours 30 minutes. Sample-type recovery (n=5) is reported at averages of 91% for serum (range 83–100%), 96% for EDTA plasma (range 90–103%), and 94% for heparin plasma (range 90–99%). Linearity data across 1:2, 1:4, 1:8, and 1:16 dilutions are provided for serum, EDTA plasma, and heparin plasma. Kit components include a pre-coated microplate (12 x 8 well strips), lyophilized standard (2 vials), Detection Solution A (120 μl), Detection Solution B (120 μl), 30X Wash Buffer (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml), and plate sealers (5 adhesive strips), stored at +4°C or -20°C as instructed.

The assay procedure involves sequential incubations at 37°C with washes between stages, the addition of 100 μl of working Detection Solution A and B per well, equilibration of TMB Substrate for 30 minutes at 37°C, addition of 90 μl of TMB Substrate per well, and termination with 50 μl of Stop Solution. The product is intended for research use only.