Rat HSD3B1/3 beta-hydroxysteroid dehydrogenase ELISA Kit

The Assay Genie kit is a Sandwich ELISA designed for the quantitative measurement of HSD3B1 in rat samples. HSD3B1 (3-beta-hydroxysteroid dehydrogenase) is a key enzyme involved in steroid hormone synthesis, playing a crucial role in the biosynthesis of steroid hormones such as estrogen, testosterone, and cortisol. Dysregulation of HSD3B1 expression has been linked to various hormonal disorders and conditions, making it a valuable biomarker for studying these diseases and potential therapeutic interventions. The research area is metabolism and the associated UniProt entry is P22071.

Using a double-antibody sandwich format with a colorimetric detection method, the kit is reactive with rat samples. Acceptable sample types include serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The assay has a reported sensitivity of 46.875 pg/mL and a working range of 78.125–5000 pg/mL. Kits should be stored at 2–8°C for 12 months.

Linearity was evaluated in serum, EDTA plasma, and heparin plasma across dilution factors of 1:2, 1:4, and 1:8, with reported ranges of 90–105%, 82–99%, and 82–100% for serum (n = 5); 90–105%, 88–101%, and 81–98% for EDTA plasma (n = 5); and 88–96%, 85–99%, and 80–100% for heparin plasma (n = 5). Recovery was tested in the same matrices, with average recovery values of 96% for serum, 95% for EDTA plasma, and 96% for heparin plasma, and corresponding recovery ranges of 88–104%, 91–100%, and 88–99%. The protocol follows a standard sandwich ELISA workflow: equilibrate reagents and TMB substrate to room temperature and set up standards, samples, and zero controls on the pre-coated plate; incubate at 37°C for 90 minutes for antigen binding; add biotin-labeled detection antibody and incubate at 37°C for 60 minutes; add HRP-Streptavidin (SABC) and incubate at 37°C for 30 minutes; develop with TMB substrate in the dark for 10–20 minutes; stop the reaction and read absorbance at 450 nm immediately.