Erba Mannheim Calcium Assay Kit

Protocol Overview

My research work involves the use of serum samples. So, prior to dissection blood samples were collected from the orbital sinus of the male Wistar rats and blood samples were allowed to clot in conical centrifuge for an hour and supernatant (serum samples) were separated using pipette. They were also centrifuged at 3000rpm for 15 mins to remove any red blood contamination. Then, as directed in the kit different reagents viz. reagent 1 and reagent 2 were allowed to attain room temperature and working reagent was prepared by mixing equal quantities of reagent 1 and reagent 2. Accordingly, 1000 µl of working reagent, 10 µl of standard/test samples was added and volume of blank was adjusted with distilled water. The samples were then incubated at 37 degrees for 1 minute and the absorbance was measured using spectrophotometer at 578nm.