Cytokine array kits

Cytokine arrays were performed from corneal tissues (n = 4–5) 3 days after SM exposure for 7 min. Lysates were prepared from the SM-exposed tissues and their respective controls; protein estimation was performed using the BioRad DC protein assay kit, following the manual instructions. The lysates were then quantified for cytokine levels using cytokine array kits (RayBiotech, Norcross, GA) according to the recommended protocol. Briefly, diluent was used to block the array epitopes. After blocking, the epitopes were incubated with the lysates. The signal detection was performed using a followed biotinylated antibody and streptavidin conjugated to a Cy3 equivalent dye. Thereafter, slides were scanned, and the data extraction was performed using RayBiotech scanning and data extraction services. RayBiotech array specific data analysis software was used for analysis and results were expressed as relative fluorescence units.