Rat Prolyl 4-hydroxylase subunit alpha-1/P4HA1 ELISA Kit

This Sandwich ELISA kit from Assay Genie is designed for the quantitative measurement of Prolyl 4-hydroxylase subunit alpha-1 (P4HA1) in rat samples using a double-antibody, colorimetric detection format. The assay is validated for use with serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples.

P4HA1 catalyses the hydroxylation of proline residues required for collagen stability, and dysregulation of P4HA1 has been implicated in fibrosis, cancer, and connective tissue disorders, highlighting its relevance as a potential biomarker and therapeutic target. The assay has a stated sensitivity of 0.188 ng/mL with a detection range of 0.313–20 ng/mL. Spiked-recovery averages are 96% for serum (range 87–104), 98% for EDTA plasma (range 89–104), and 93% for heparin plasma (range 87–103), each based on n = 5. Reported linearity at 1:2, 1:4, and 1:8 dilutions is 90–103%, 84–97%, and 82–97% for serum; 90–99%, 85–99%, and 89–94% for EDTA plasma; and 85–102%, 82–96%, and 89–95% for heparin plasma. Reagents are stored at 2–8°C for up to 12 months.

The recommended workflow involves equilibrating reagents and the TMB substrate to room temperature, loading standards, samples and blanks for a 90-minute primary incubation at 37 °C, adding biotin-labelled detection antibody for 60 minutes at 37 °C, incubating with HRP-Streptavidin (SABC) for 30 minutes at 37 °C, developing colour with TMB substrate in the dark for 10–20 minutes, and reading absorbance at 450 nm immediately after adding stop solution.