Human Phenylalanine Hydroxylase ELISA Kit

Offered under the Antibodies.com brand, this sandwich ELISA (sELISA) is designed for the in vitro quantitative determination of human Phenylalanine Hydroxylase (PAH) in tissue homogenates, cell lysates, or other biological fluids. The microplate is pre-coated with an antibody specific for PAH; standards and test samples are added so that PAH is captured by the immobilised antibody. After washing, a Biotinylated Anti-PAH Antibody binds the captured PAH, followed by an HRP-Avidin conjugate. TMB substrate is then added and catalysed to produce a blue-coloured product, which turns yellow upon addition of the acidic stop solution; absorbance is read at 450 nm in a colorimetric microplate reader, and PAH concentration is calculated by reference to the standard curve.

The quantitative sandwich assay has a reported range of 1.56–100 ng/ml and a sensitivity of 0.56 ng/ml, with a total assay time of 4h 30m. Recovery has been validated in serum (80–102%, average 91%, n=5), EDTA plasma (81–100%, average 90%, n=5) and heparin plasma (80–89%, average 84%, n=5). Dilution linearity across 1:2, 1:4, 1:8 and 1:16 dilutions has also been demonstrated for serum (74–107%), EDTA plasma (79–108%) and heparin plasma (82–105%). Kit components should be stored at +4°C or –20°C as indicated, and the kit must not be used past its expiration date.