Mouse Glycogen Phosphorylase, Liver ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is intended for the in vitro quantitative determination of mouse PYGL (Glycogen Phosphorylase, Liver) in serum, plasma, tissue homogenates or other biological fluids. Reactivity is mouse-specific and the assay is performed on a pre-coated 96-well microplate arranged as 12 x 8 well strips, with detection carried out on a colorimetric microplate reader.

The kit uses the sandwich enzyme immunoassay technique. An antibody specific for PYGL is pre-coated onto the plate; standards and samples are added and PYGL is captured by the immobilized antibody. Following incubation and washing, Biotinylated Anti-PYGL Antibody is added, followed by HRP-Avidin conjugate. TMB substrate is then used to visualize the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of PYGL captured; absorbance is read at 450 nm and the concentration is calculated by reference to a standard curve. Total assay time is given as 4 h 30 min.

Reported performance figures include a sensitivity of <0.053 ng/ml and an assay range of 0.156–10 ng/ml. Recovery values were: serum (n=5) range 80%–102%, average 91%; EDTA plasma (n=5) range 81%–100%, average 90%; heparin plasma (n=5) range 80%–89%, average 84%. Dilution linearity data for 1:2, 1:4, 1:8 and 1:16 dilutions were: serum 87–91%, 87–107%, 74–101% and 92–97%; EDTA plasma 90–105%, 84–101%, 90–101% and 79–108%; heparin plasma 84–95%, 92–105%, 82–105% and 89–91%. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past the expiration date.