Human ZP3/Zona pellucida sperm-binding protein 3 ELISA Kit

Assay Genie supplies a Sandwich ELISA, Double Antibody format kit for the quantitative detection of human Zona Pellucida Sperm-Binding Protein 3 (ZP3). The assay is designed for human reactivity and accepts serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid sample types, with detection carried out by a colorimetric ELISA method. The kit reports a sensitivity of 0.375 ng/mL and a detection range of 0.625-40 ng/mL, and is stored at 2-8°C for 12 months.

Assay performance characteristics include linearity tested at 1:2, 1:4 and 1:8 dilutions: serum (n=5) 89-99% 85-97% 91-104%, EDTA plasma (n=5) 83-100% 83-94% 86-100%, and heparin plasma (n=5) 81-100% 80-100% 84-97%. Spike-and-recovery values are serum (n=5) range 87-104% average 93%, EDTA plasma (n=5) range 86-104% average 91%, and heparin plasma (n=5) range 86-103% average 97%.

The procedure runs as a six-step sandwich ELISA: equilibrate reagents and TMB substrate and load standards, samples and a zero control 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 for 10-20 minutes in the dark; then add stop solution and read absorbance at 450 nm. Sample preparation covers serum (clot, 1000 × g, 20 minutes), plasma (anticoagulant tube, 1000 × g, 15 minutes, 2-8°C), tissue homogenate (PBS with protease inhibitors), cell culture supernatant (2500 rpm, 5 minutes), and cell lysate (lysis buffer with protease inhibitors). Kit components include a dismountable ELISA Microplate, lyophilized standard, 100X biotin-labeled antibody, 100X HRP-Streptavidin conjugate (SABC), TMB substrate, sample dilution buffer, antibody dilution buffer, SABC dilution buffer, stop solution, 25X wash buffer, plate sealer and a technical manual.