Human N-Acylsphingosine Amidohydrolase 1 ELISA Kit

A sandwich enzyme-linked immunosorbent assay (ELISA) designed for the in vitro quantitative determination of human N-Acylsphingosine Amidohydrolase 1 (ASAH1) in tissue homogenates, cell lysates, or other biological fluids. Distributed by Antibodies.com, the kit employs a pre-coated 96-well microplate in a 12 x 8 well strip format with colorimetric detection read at 450 nm in a microplate reader.

The assay has a reported sensitivity of less than 0.61 ng/ml and a measuring range of 1.56–100 ng/ml, with a total run time of approximately 4 hours 30 minutes. The procedure uses a biotinylated anti-ASAH1 antibody and an HRP-avidin conjugate for detection, with TMB substrate producing a yellow colour following addition of acidic stop solution.

Spike-and-recovery performance was evaluated in three matrices: serum (n=5) showed an average of 91% across a range of 80–102%; EDTA plasma (n=5) averaged 90% across 81–100%; and heparin plasma (n=5) averaged 84% across 80–89%. Dilution linearity was also assessed at 1:2, 1:4, 1:8, and 1:16 in each matrix. Components supplied include the pre-coated microplate, lyophilized standard, detection solutions A and B, wash buffer, sample dilution buffer, TMB substrate, stop solution, and plate sealers. Components should be stored at +4°C or -20°C as indicated, and the kit should not be used past its expiration date.