Kc junior microplate reader software

Insulin concentrations in each specimen (culture medium and cell extracts) were measured using Ultra Sensitive Human Insulin ELISA Kit (Milllipore, EZHI-14K) according to the vendor's instructions and insulin concentrations were calculated using KC Junior^® microplate reader software (Bio-Tek Instruments, Inc.) [31 (link)]. Briefly, insulin standards and appropriately diluted samples were added to an insulin antibody-coated 96-well microplate and, incubated for 1 hour at room temperature after adding the detection antibody. After washing five times, anti-human insulin enzyme conjugate was added to the well and incubated for 30 min at room temperature. After washing five times, an enzyme substrate solution was added and then incubated 10 to 15 min at RT in the dark. The reaction was halted by adding 1 N sulfuric acid. Absorbance at 450 nm was read with a μQuant microplate reader (Bio-Tek Instruments, Inc., Winooski, VT) and concentrations were calculated by KC Junior® microplate reader software (Bio-Tek Instruments, Inc.) [17 (link)].