An Easy Way To Quantify Thrombin-Antithrombin Complexes Using Abcam ELISA Kit

Protocol Overview

Ocular Blood was collected in a heparinized tube from mice of all groups and mixed gently. Tubes containing heparinized blood was centrifuged at 275 xg for 5 minutes and pale yellow supernatant (Plasma) was collected. Plasma was used for quantification of Thrombin-Antithrombin Complexes ELISA as per manufacture instructions. In brief, plasma was added to ELISA plate well and incubated for 2 hours at room temperature. After washing wells were washed five times with washing buffer and biotin conjugated anti- Thrombin-Antithrombin Complexes antibody was added and incubated for 1 hour at room temperature. Wells were washed four times and treated with streptavidin peroxidase conjugate. Finally, colour was developed by adding chromogen substrate and colour intensity was read under a microplate reader.