Human Proteasome Subunit Beta Type 6 ELISA Kit

From Antibodies.com, this sandwich ELISA kit is designed for the in vitro quantitative determination of human Proteasome 20S beta 6 in tissue homogenates, cell lysates, or other biological fluids. It is reactive with human samples and uses a pre-coated microplate format comprising 12 x 8 well strips read at 450 nm on a colorimetric microplate reader.

The assay uses the sandwich enzyme immunoassay technique. Standards and test samples are added to wells pre-coated with an antibody specific for Proteasome 20S beta 6, where the target is captured by the immobilised antibody. After washing, a Biotinylated Anti-Proteasome 20S beta 6 Antibody is added and binds the captured target. An HRP-Avidin conjugate is then added, and following incubation and washing, TMB substrate solution is used to visualise the HRP enzymatic reaction, generating a blue colour that changes to yellow upon addition of acidic stop solution. The intensity of yellow is proportional to the amount of Proteasome 20S beta 6 in each well, and the concentration is calculated by referring absorbance readings to a standard curve.

Reported performance characteristics include a sensitivity of 0.053 ng/ml, an assay range of 0.156-10 ng/ml, and a total assay time of 4 hours 30 minutes. Sample recovery has been validated in serum (average 91%, range 83-99%, n=5), EDTA plasma (average 89%, range 80-99%, n=5), and heparin plasma (average 90%, range 82-99%, n=5), with linearity demonstrated across dilution factors of 1:2, 1:4, 1:8, and 1:16. Kit components are stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.