Rat Nephrin ELISA Kit

Designed for the precise measurement of nephrin levels in rat samples, this colorimetric Sandwich ELISA from Assay Genie supports research into the kidney filtration barrier and associated conditions such as diabetic nephropathy and glomerulonephritis. The assay is validated for serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid sample types, and falls within the cell biology research area.

The kit is stored at 2–8°C for 12 months. The procedure uses a pre-coated plate: equilibrate reagents and TMB substrate to room temperature; incubate standards, samples and controls at 37°C for 90 minutes for antigen binding; add biotin-labelled 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; then add stop solution and read absorbance at 450 nm.

Linearity across dilution ratios of 1:2, 1:4 and 1:8 (n=5) was 87–101%, 85–98% and 81–95% for serum; 87–98%, 85–97% and 86–97% for EDTA plasma; and 82–100%, 80–89% and 80–100% for heparin plasma. Spike-and-recovery ranges with average values were 85–105% (average 98%) for serum, 87–99% (average 93%) for EDTA plasma, and 86–100% (average 95%) for heparin plasma.