Green Fluorescent Cathepsin B Assay

• Description:

  The quenched substrate Rhodamine 110-(RR)2, fluoresces green upon cleavage by active cathepsins, allowing the monitoring of intracellular cathepsin activity in vitro using live whole cells. Analyze using flow cytometry or fluorescence microscopy.

• Assay Principle:

  1. Dilute 10X Cellular Assay Buffer 1:10 with diH2O., 2. Prepare samples and controls in fresh cell culture medium or 1X Cellular Assay Buffer., 3. Reconstitute R110-(RR)2 with 50 µL DMSO., 4. Dilute R110-(RR)2 1:5 by adding 200 µL PBS., 5. Add diluted R110-(RR)2 to each sample at 1:50 (e.g., add 10 µL of diluted R110-(RR)2 to 490 µL of sample)., 6. Incubate while protected from light., 7. If desired, label with additional stains, such as Hoechst 33342, DAPI, or an antibody., 8. Analyze with a flow cytometer or fluorescence microscope. R110-(RR)2 excites at 500 nm and emits at 525 nm.

• Shipping Conditions:

  Blue Ice; Dry Ice

• Storage Conditions:

  4°C;-20°C