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Normal (left) and keratoconus
(right) corneal fibroblasts were treated with 200uM H2O2 for 1hr, washed, and allowed to recover for 1-3hrs. The culture media was removed and replaced with 1x
FAM-FLICA™ Caspases 3&7 (kit cat.# 93) solution (in culture media) at 300ul/well for 1hr. The cell layer was washed 3 times with 1x wash buffer; 300ul wash buffer was added to keep the cells from drying. Keratoconus corneal fibroblasts treated with H2O2
(right) show a significant increase in caspases 3&7 activity compared to normal cells
(left). Non-apoptotic cells are dark in background. Data
courtesy of Dr. Cristina
Kenney, M.D., Ph.D. Dept. of Ophthalmology,
UC Irvine.
FLICA™ is a cell-permeant fluorescent probe used to quantitate caspase activity in
cultured cells. It is a direct stain: once labeled, tissues are ready
for analysis and no additional staining steps are necessary- just wash and
read. FLICA™ is very specific. Only active caspase enzymes will covalently bind with the
reagent, therefore only cells undergoing apoptosis will fluoresce. FLICA™ is very easy to use. Just
add it to the cell culture media, let it incubate 1-4 hours,
wash, and analyze with a fluorescence microscope, plate
reader, or flow cytometer. Dying apoptotic tumor
cells fluoresce
green
using FAM-FLICA™
or red using SR-FLICA™.
Try a kit today!
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