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CS0350

Sigma-Aldrich

Cathepsin Detection Assay

Cathepsin L Detection Assay, sufficient for 100 assays

Pricing and availability is not currently available.

usage

sufficient for 100 assays

storage temp.

2-8°C

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Biochem/physiol Actions

Cathepsins are apoptosis markers associated with Alzheimer′s disease, numerous types of cancer, autoimmune arthritis, and the breakdown of bone structure seen with osteoporosis.

Principle

Cathepsin B, K, and L protease activities can be detected within whole living cells using substrate-based fluorescent assays, which utilize cathepsin target sequence peptides, derivatives of the cresyl violet fluorophore, conjugated to Magic Red™ (MR) fluorogenic substrate (cresyl violet). Following enzymatic cleavage inside the living cells at one or both arginine (R) amide linkage sites, the mono and non-substituted cresyl violet fluorophores generate red fluorescence when excited at 550-590 nm.
Cathepsin L detection assay detects protease activity within whole living cells utilizing fluorogenic substrate (z-Phenylalanine -Arginine)2(z-FR)2 derivative of the cresyl violet fluorophore, which easily penetrates the cell membrane and the membranes of the internal cellular organelles. Hoechst stain is used to label the cell nuclei after labeling with the MR- Cathepsin L substrate. It is visualized under a microscope using a UV-filter with excitation at 365 nm and emission at 480 nm. Acridine orange (AO) helps identify lysosomes and other intracellular organelles. In the acidic pH of the lysosome AO molecules aggregate. Aggregated AO fluoreses orange rather than green thus clearly differentiating the lysosomes from the other organelles.

Legal Information

Magic Red™ trademark is owned by MP Biomedicals (Formerly ESP).

Kit Components Only

Product No.
Description

  • MR-(FR)2 substrate 1 mL/vial

  • Hoechst 33342 stain 1 mL

  • Acridine orange stain (AO) .5 mL

Certificate of Analysis
Certificate of Origin
Jörg Stypmann et al.
Proceedings of the National Academy of Sciences of the United States of America, 99(9), 6234-6239 (2002-04-25)
Dilated cardiomyopathy is a frequent cause of heart failure and is associated with high mortality. Progressive remodeling of the myocardium leads to increased dimensions of heart chambers. The role of intracellular proteolysis in the progressive remodeling that underlies dilated cardiomyopathy...

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