Human Protein Kinase, AMP Activated Beta 1 ELISA Kit

Antibodies.com provides a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) for the in vitro quantitative determination of human AMPK beta 1 in tissue homogenates or other biological fluids. The assay uses a 96-well microtiter plate pre-coated with an antibody specific for AMPK beta 1. Standards and test samples are added to the wells where the target binds to the immobilized antibody, followed by washing and incubation with a Biotinylated Anti-AMPK beta 1 Antibody. After further washing, HRP-Avidin conjugate is added and the plate is incubated. TMB substrate solution then produces a blue-coloured product that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of AMPK beta 1 captured in each well, and concentration is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

The assay range is 0.312-20 ng/ml with a sensitivity of 0.124 ng/ml, run on a colorimetric microplate reader. The total assay time is 4 hours 30 minutes. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date. Average recovery was 91% for serum, 90% for EDTA plasma, and 90% for heparin plasma, with dilution linearity demonstrated at 1:2, 1:4, 1:8, and 1:16 dilutions across each sample type.