Human PRNP/CD230/Prion ELISA Kit

A colorimetric sandwich ELISA from Assay Genie for the quantitative detection of human PRNP (CD230), the prion protein encoded by the PRNP gene that is implicated in prion diseases such as Creutzfeldt-Jakob disease. The double-antibody sandwich assay is validated for use with serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples.

Stated analytical performance includes a sensitivity of 0.188 ng/mL and an assay range of 0.313–20 ng/mL. Linearity assessed by serial dilution (n=5) was 93–104%, 88–99% and 86–101% in serum at 1:2, 1:4 and 1:8; 86–101%, 83–99% and 82–97% in EDTA plasma; and 83–92%, 81–98% and 81–95% in heparin plasma. Spike-and-recovery ranged from 85–100% in serum (average 90%), 88–101% in EDTA plasma (average 93%) and 86–101% in heparin plasma (average 91%).

The procedure involves equilibrating reagents and TMB substrate to room temperature, loading standards, samples and blanks onto the pre-coated plate and incubating at 37°C for 90 minutes, followed by incubation with biotinylated detection antibody (37°C, 60 min), HRP-streptavidin SABC (37°C, 30 min), TMB colour development in the dark for 10–20 minutes, and absorbance reading at 450 nm after adding stop solution. Kit components include a dismountable ELISA microplate, lyophilized standard, biotin-labeled antibody, HRP-streptavidin conjugate (SABC), TMB substrate, sample dilution buffer, antibody dilution buffer, SABC dilution buffer, stop solution, wash buffer, plate sealers and a technical manual. Unopened kits are stored at 2–8°C for 12 months.