Jun 25, 2024
Magic Red Cathepsin B activity assay
- Razaul Karim1
- 1University of Minnesota, Team Lee
Protocol Citation: Razaul Karim 2024. Magic Red Cathepsin B activity assay. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov192b2lr2/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 07, 2024
Last Modified: June 25, 2024
Protocol Integer ID: 102393
Keywords: Cathepsin B , scanning microscope, signal intensity
Funders Acknowledgement:
ASAP
Grant ID: 000592
Abstract
This protocol details the magic red Cathepsin B activity assay.
Materials
Assay Kits:
- Magic Red Fluorescent Cathepsin B assay kit (Immunochemistry Technologies, CA).
Coverslips:
- Corning® BioCoat™ Poly-D-Lysine #1 German Glass CoverslipMerckCatalog #354086
Cell Culture/Treatments
Cell Culture/Treatments
15m
0.5x10^4/ml of cells plated on coverslips.
PFF-488 treatment.
PBS/trypsin (0.01%) wash.
1x Magic Red solution for last 00:15:00 at 37 °C in cell culture incubator.
15m
Imaging
Imaging
Live-Cell imaging.
Leica Stellaries 8 confocal laser scanning microscope, assembled with live cell chamber with CO2 gas, temperature and humidity controlled.
Image analysis:
Image J. quantification of the % area of the cell covered by Magic Red signal, as well as total signal intensity per cell.