Canine High Mobility Group Protein 1 ELISA Kit

From Antibodies.com, this Sandwich ELISA is intended for the in vitro quantitative determination of canine HMGB1 (High Mobility Group Protein 1) in serum, plasma or other biological fluids. The kit uses a pre-coated microplate and follows the sandwich enzyme immunoassay technique: standards and samples are added to the wells where HMGB1 is captured by the immobilised antibody, followed by incubation with a Biotinylated Anti-HMGB1 Antibody, HRP-Avidin conjugate and TMB substrate. The resulting blue product changes to yellow on addition of acidic stop solution, and the intensity of yellow is proportional to the amount of HMGB1 captured in each well.

Absorbance is read at 450 nm on a colorimetric microplate reader and the HMGB1 concentration is calculated by reference to a standard curve. Manufacturer-reported sensitivity is less than 1.24 ng/ml, with an assay range of 3.12–200 ng/ml, and a total assay time of 4 hours 30 minutes.

The manufacturer reports recovery across five samples per matrix, with averages of 91% for serum (range 80–102%), 90% for EDTA plasma (range 81–100%) and 84% for heparin plasma (range 80–89%). Linearity is provided for 1:2, 1:4, 1:8 and 1:16 dilutions across serum, EDTA plasma and heparin plasma. Kit components are stored at +4°C or −20°C as instructed, and the kit should not be used past its expiration date.