Ph-F Cholinesterase Detection Reagent
Detect cholinesterase activity in vitro using ICT's inhibitor Ph-F
With ICT’s inhibitor probe Ph-F, researchers have an easy way to directly quantify and localize active cholinesterase enzymes in live, intact cells - no lysis or permeabilization steps are required.
The probe uses physostigmine, a known cholinesterase inhibitor, linked to a green fluorescent label for cholinesterase detection. It is cell permeant, so there is no need to lyse the cells or permeabilize the membranes.
More Information:
- Read a paper on cholinesterase
- Read the Ph-F probe brochure
- Read the Ph-F probe manual
Ph-F Cholinesterase Detection Reagent
Localization of cholinesterase with Ph-Fl in the nerve-muscle junctions (end-plates) in C57 mouse diaphragm muscle tissue. The diaphragm muscle was dissected and fragments of it incubated in PBS containing 20μM Ph-Fl for 1 hour. The muscle was then rinsed for 20 minutes in PBS, stretched on a microscope slide, mounted under a coverslip, and examined by fluorescence microscopy using blue light excitation. Notice the retention of the fluorochrome in the structures with typical features of end-plates (arrows).
Sample protocol:
- Culture your cells.
- Subject cells to your experimental protocol.
- Reconstitute the reagent to form the stock concentrate.
- Dilute the stock concentrate to the final working solution.
- Add the working solution directly to the cell culture for labeling.
- If desired, concurrently label cells with DAPI, or SR-VAD-FMK.
- Incubate for 1-4 hours.
- Wash cells with culture media.
- If desired, fix cells.
- Analyze data with a fluorescence microscope, fluorescence plate reader, or flow cytometer.