Mouse Adrenergic Receptor Beta 3 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com designed for the in vitro quantitative determination of mouse beta 3 Adrenergic Receptor in tissue homogenates or other biological fluids. The assay uses a pre-coated 96-well microplate configured as 12 x 8 well strips and employs the sandwich enzyme immunoassay technique with a biotinylated detection antibody and HRP-Avidin conjugate. Signal is visualised using TMB substrate, with the enzymatic reaction producing a blue product that turns yellow upon addition of acidic stop solution; absorbance is read at 450 nm in a microplate reader and the concentration of beta 3 Adrenergic Receptor is calculated by reference to a standard curve.

The standard assay range is 15.6-1000 pg/ml, with a sensitivity of <5.9 pg/ml. The total assay time is approximately 4 hours 30 minutes. Kit components include a Pre-Coated 96 Well Microplate (12 x 8 Well Strips), Lyophilized 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 5 Adhesive Plate Sealers. Components are stored at +4°C or -20°C as instructed and the kit should not be used past its expiration date.

For standard preparation, the lyophilized standard is reconstituted in 1 ml of Sample Dilution Buffer to yield a 1000 pg/ml stock solution, which is then used to prepare a 1:2 serial dilution series producing 500, 250, 125, 62.5, 31.3, and 15.6 pg/ml standards. Detection Solutions A and B are diluted 1:100 in Sample Dilution Buffer before use. The procedure involves sequential incubations at 37°C with washing steps between additions of Detection Solution A and Detection Solution B, followed by TMB substrate incubation in the dark for 15-25 minutes and Stop Solution addition to terminate the reaction.