Rat RNase3/ECP ELISA Kit

From Assay Genie, this Sandwich ELISA uses a double-antibody, colorimetric format to quantify rat RNase3 (Eosinophil Cationic Protein; synonyms ECP, Ribonuclease A3; UniProt P70709) in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Reactivity is rat-specific.

Sensitivity is 37.5 pg/mL and the standard range is 62.5-4000 pg/mL. Linearity at 1:2, 1:4, and 1:8 dilutions: serum (n=5) 85-102%, 83-94%, 87-99%; EDTA plasma (n=5) 89-105%, 87-100%, 80-92%; heparin plasma (n=5) 87-105%, 86-94%, 82-94%. Recovery: serum (n=5) 88-100% (average 94%); EDTA plasma (n=5) 85-104% (average 99%); heparin plasma (n=5) 90-104% (average 96%). The kit is stored at 2-8°C for 12 months; microplate strips and lyophilized standard are also stable at -20°C for 12 months.

The assay procedure: equilibrate reagents and TMB substrate at room temperature; load standards, samples and blanks and incubate 90 minutes at 37°C; add biotinylated detection antibody and incubate 60 minutes at 37°C; add HRP-Streptavidin (SABC) and incubate 30 minutes at 37°C; add TMB substrate and incubate 10-20 minutes in the dark; add stop solution and read absorbance at 450 nm. Sample preparation includes serum clotting (centrifuge 1000 × g, 20 minutes), plasma collection with anticoagulant (centrifuge 1000 × g, 15 minutes, 2-8°C), tissue homogenization in PBS with protease inhibitors, cell culture supernatant clarification (2500 rpm, 5 minutes), and cell lysis with buffer containing protease inhibitors.