Reagents and instruments for immunology, cell biology and molecular biology.

Magic Red™ Cathepsin K BioAssay™ Kit

Print

Cat# 474031-25T

Size : 25Tests

Brand : US Biological

Request more information

474031 Magic Red™ Cathepsin K BioAssay™ Kit

Clone Type

Polyclonal

Shipping Temp

Blue Ice

Storage Temp

4°C/-20°C

Sample Type:|Cell culture||Reagent Name: |MR-LR2||Target:|Cathepsin K||Excitation/Emission:|592nm/628nm||Method of Analysis:|Fluorescent Plate Reader, Fluorescence Microscope||Intended Use:|Quantitate and monitor intracellular cathepsin K activity over time in vitro. The Magic Red reagent in this BioAssay™ fluoresces red upon cleavage by active cathepsin enzymes. Analyze the fluorescent signal using fluorescence microscopy or a fluorescent plate reader.||Test Principle:|Elevated cathepsin enzyme activity in serum or the extracellular matrix can signify a number of pathological conditions. Magic Red BioAssay™ kits are used by researchers seeking to quantitate and monitor cathepsin activity in cultured cells and tissues. This Magic Red BioAssay™ kit detects cathepsin K activity.||The Magic Red reagent MR-LR2 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.||Kit Components:|25 tests:|Magic Red Reagent (MR-LR2), 1 25-test vial|Hoechst 33342 Stain, 1x1mL|Acridine Orange Stain, 1x0.5mL||100 tests:|Magic Red Reagent (MR-LR2), 1 100-test vial|Hoechst 33342 Stain, 1x1mL|Acridine Orange Stain, 1x0.5mL||Storage and Stability:|Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Applications

Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. Toxicity and Hazards: All products should be handled by qualified personnel only, trained in laboratory procedures.

References

1. Buhling, F. et al. Review: novel cysteine proteases of the papain family. Adv Exp Med Biol 477, 241-254, doi:10.1007/0-306-46826-3_26 (2000).|2. Gerber, A., Welte, T., Ansorge, S. & Buhling, F. Expression of cathepsins B and L in human lung epithelial cells is regulated by cytokines. Adv Exp Med Biol 477, 287-292, doi:10.1007/0-306-46826-3_31 (2000).|3. Bank, U., Kruger, S., Langner, J. & Roessner, A. Review: peptidases and peptidase inhibitors in the pathogenesis of diseases. Disturbances in the ubiquitin-mediated proteolytic system. Protease-antiprotease imbalance in|inflammatory reactions. Role of cathepsins in tumour progression. Adv Exp Med Biol 477, 349-378, doi:10.1007/0-306-46826-3_38 (2000).|4. Frosch, B. A., Berquin, I., Emmert-Buck, M. R., Moin, K. & Sloane, B. F. Molecular regulation, membrane association and secretion of tumor cathepsin B. APMIS 107, 28-37(1999).|5. Guinec, N., Dalet-Fumeron, V. & Pagano, M. “In vitro” study of basement membrane degradation by the cysteine proteinases, cathepsins B, B-like and L. Digestion of collagen IV, laminin, fibronectin, and release of gelatinase activities|from basement membrane fibronectin. Biol Chem Hoppe Seyler 374, 1135-1146 (1993).|6. Kos, J., Werle, B., Lah, T. & Brunner, N. Cysteine protein-ases and their inhibitors in extracellular fluids: markers for diagnosis and prognosis in cancer. Int J Biol Markers 15, 84-89 (2000).|7. Buhling, F. et al. Cathepsin K expression in human lung.|Adv Exp Med Biol 477, 281-286, doi:10.1007/0-306-46826-|3_30 (2000).|8. Bossard, M. J. et al. Proteolytic activity of human osteoclast cathepsin K. Expression, purification, activation, and substrate identification. J Biol Chem 271, 12517-12524 (1996).|9. Leung-Toung, R., Li, W., Tam, T. F. & Karimian, K. Thiol-dependent enzymes and their inhibitors: a review. Curr Med Chem 9, 979-1002 (2002).|10. Van Noorden, C. J. et al. Ala-Pro-cresyl violet, a synthetic fluorogenic substrate for the analysis of kinetic parameters of dipeptidyl peptidase IV (CD26) in individual living rat hepatocytes. Anal Biochem 252, 71-77, doi:10.1006/|abio.1997.2312 (1997).|11. Turk, B., Turk, D. & Turk, V. Lysosomal cysteine proteases: more than scavengers. Biochim Biophys Acta 1477, 98-111 (2000).|12. Boonacker, E. & Van Noorden, C. J. Enzyme cytochemical techniques for metabolic mapping in living cells, with special reference to proteolysis. J Histochem Cytochem 49, 1473-1486, doi:10.1177/002215540104901201 (2001).