Magic Red Cathepsin L Assay Kit

$185.00$494.00

Quantitate and monitor intracellular cathepsin L activity over time in vitro. The Magic Red reagent in this assay fluoresces red upon cleavage by active cathepsin enzymes. Analyze the fluorescent signal using fluorescence microscopy or a fluorescent plate reader.

Catalog # Size Quantity Price
941 25 tests $185.00
942 100 tests $494.00
Catalog #: 941, 942 Categories: , ,

Elevated cathepsin enzyme activity in serum or the extracellular matrix can signify a number of pathological conditions. ICT’s Magic Red assay kits are used by researchers seeking to quantitate and monitor cathepsin activity in cultured cells and tissues. This Magic Red kit detects cathepsin L

The Magic Red reagent MR-FR2 enters each cell in a non-fluorescent state. If cathepsin enzymes are active, the Magic Red substrate is cleaved and the cresyl violet fluorophone will become fluorescent upon excitation. As enzyme activity progresses and more Magic Red substrate is cleaved, the signal will intensify, allowing researchers to watch the color develop over time. Samples can be analyzed by fluorescence microscopy or fluorescent plate reader. Samples may also be analyzed with Acridine Orange or Hoechst stain to detect lysosomal organelle structure or nuclear morphology respectively.

1. Prepare samples and controls
2. Reconstitute Magic Red by adding 50 µL DMSO.
3. Dilute Magic Red 1:10 by adding 450 µL diH2O.
4. Add 20 µL Magic Red to each sample (~ 500 µL aliquot of cultured cells).
5. Incubate while protected from light.
6. Watch color start to develop within 15 minutes.
7. If desired, label with additional stains, such as Hoechst, DAPI, Acridine orange, or an antibody.
8. If desired, fix or embed cells.
9. Analyze with a fluorescence microscope or fluorescence plate reader. Magic Red has an optimal excitation at 592 nm and emission at 628 nm.

If working with adherent cells, please see the manual for additional protocols.

Cathepsin L-positive Jurkat cells fluoresce red after staining with MR-(FR)2. Red fluorophore concentrates inside lysosomes (Dr. Brian W. Lee, ICT).mrfrcathldata

• Reagent name: MR-FR2
• Target: Cathepsin L
• Excitation/Emission: 592 nm / 628 nm
• Method of Analysis: Fluorescent Plate Reader, Fluorescence Microscope
• Types of Samples: Cell culture
• Storage: 2-8°C
• Shipping: Ships overnight (domestic), International Priority Shipping

Kit 941: 25 tests
• Magic Red Reagent (MR-FR2), 1 25-test vial, #6137
• Hoechst 33342 Stain, 1 mL, #639
• Acridine Orange Stain, 0.5 mL, #6130
• Kit Manual

Kit 942: 100 tests
• Magic Red Reagent (MR-FR2), 1 100-test vial, #6138
• Hoechst 33342 Stain, 1 mL, #639
• Acridine Orange Stain, 0.5 mL, #6130
• Kit Manual

Matsumoto A, Pasut A, Matsumoto M, Yamashita R, Fung J, Monteleone E, Saghatelian A, Nakayama KI, Clohessy JG, Pandolfi PP. mTORC1 and muscle regeneration are regulated by the LINC00961-encoded SPAR polypeptide. Nature. 541(7636): 228-232. Abstract.

“Cathepsin K and L activity was monitored with MagicRed Cathepsin Assay Kit (ImmunoChemistry Technologies) according to the manufacturer’s recommendations.”

Marwaha R, Arya SB, Jagga D, Kaur H, Tuli A, Sharma M. The Rab7 effector PLEKHM1 binds ArI8b to promote cargo traffic to lysosomes. J. Cell. Biol. 2017. 216(4): 1051-1070. Full text.

Kang HT, Park JT, Choi K, Kim Y, Choi HJC, Jung CW, Lee YS, Chul Park S. Chemical screening identifies ATM as a target for alleviating senescence. Nat Chem Biol. 2017. Jun;13(6):616-623. Epub 2017 Mar 27. Abstract.

“Cathepsin activity was determined with MagicRed cathepsin B (CTSB) and cathepsin L (CTSL) assay kits (MR-CTSB and MR-CTSL, respectively; ImmunoChemistry Technologies), according to the manufacturer’s instructions.”

“The Magic Red Cathepsin L Assay kit to monitor activity of the pH-sensitive protease cathepsin L was purchased from ImmunoChemistry Technologies.”

Guo C, Zhu Z, Guo Y, Wang X, Yu P, Xiao S, Chen Y, Cao Y, and Liu X. Heparanase upregulation contributes to porcine reproductive and respiratory syndrome virus release. J. Virol. 2017. May 10. pii: JVI.00625-17. doi: 10.1128/JVI.00625-17. [Epub ahead of print]. Abstract.

” Enzymatic activity assay Cathepsin L activity in living cells was monitored using the Magic Red Cathepsin L detection kit (ImmunoChemistry Technologies, MN, USA). Briefly, cells were seeded onto coverslips, and then inoculated with PRRSV at an multiplicity of infection (MOI) of 1, 2, or 3. After incubation for the indicated times, Magic Red staining solution which is a non-cytotoxic substrate that fluoresces red upon cleavage by active cathepsins was added for 1 h at 37 °C. ”