Company with Director Field Operations jobs. Christiana Care Health System. We are of the country's largest health care providers, ranking 21st in the nation for hospital admissions and 100 best hospitals. Jobs (271) Reviews (343) Photos (2) Salaries (183) Director of Operations salaries in United States. Detect active caspase enzymes with the FAM-FLICA Poly Caspase Assay Kit. This in vitro assay employs the fluorescent inhibitor probe FAM-VAD-FMK to label active caspase enzymes in living cells. Analyze samples using fluorescence microscopy, a fluorescence plate reader, or flow cytometry.

Caspases play important roles in apoptosis and inflammation. ICT’s FLICA assay kits are used by researchers seeking to quantitate apoptosis via caspase activity in cultured cells and tissues. The FAM FLICA Poly Caspase probe allows researchers to assess caspase activation. The FLICA reagent FAM-VAD-FMK enters each cell and irreversibly binds to activated caspases. Because the FAM-VAD-FMK FLICA reagent becomes covalently coupled to the active enzymes, it is retained within the cell, while any unbound FAM-VAD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining green fluorescent signal is a direct measure of the active caspase enzyme activity present in the cell at the time the reagent was added.

Cells that contain the bound FLICA can be analyzed by a fluorescence plate reader, fluorescence microscopy or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 16 hours using the fixative included in the kit. Unfixed samples may also be analyzed with Propidium Iodide or Hoechst 33342 to detect changes in necrosis or nuclear morphology, respectively. Prepare samples and controls 2. Dilute 10X Apoptosis Wash Buffer 1:10 with diH 20.

Reconstitute FLICA with 50 µL DMSO. Dilute FLICA 1:5 by adding 200 µL PBS. Add diluted FLICA to each sample at 1:30 (e.g., add 10 µL to 290 µL of cultured cells). Incubate approximately 1 hour.

Remove media and wash cells 3 times: add 1X Apoptosis Wash Buffer and spin cells. If desired, label with additional stains, such as Hoechst, Propidium Iodide, 7-AAD, or an antibody. If desired, fix cells. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. FAM-FLICA excites at 492 nm and emits at 520 nm.

If working with adherent cells, please see the manual for additional protocols. Jurkat cells dually stained with Hoechst and poly-caspases FLICA (cat. Caspase activity is revealed by green fluorescence in the middle cell, indicating that only this cell is apoptotic. Cell at left is also dying (scattered blue), but is not apoptotic because it is not green. Cell in upper right is healthy (concentrated blue nucleus, no green FLICA). Philippine electrical code part 1 free download.