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> Magic Red™ Cathepsin B BioAssay™ Kit

Magic Red™ Cathepsin B BioAssay™ Kit

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Référence 474030-100T

Conditionnement : 100Tests

Marque : US Biological

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474030 Magic Red™ Cathepsin B BioAssay™ Kit

Clone Type
Polyclonal
Shipping Temp
Blue Ice
Storage Temp
4°C/-20°C

Sample Type:|Cell culture||Reagent Name: |MR-RR2||Target:|Cathepsin-B||Excitation/Emission:|592nm/628nm||Method of Analysis:|Fluorescence Plate Reader, Fluorescence Microscope||Intended Use:|Quantitate and monitor intracellular cathepsin-B activity over time in vitro. The Magic Red substrate in this BioAssay™ fluoresces red upon cleavage by active cathepsin enzymes. Analyze the fluorescent signal using fluorescence microscopy or a fluorescence 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 BioAssay™ kit detects cathepsin-B activity.||To use Magic Red, add the substrate directly to the cell culture media, incubate, and analyze. Because it is cell permeant, it easily penetrates the cell membrane and the membranes of the internal cellular organelles – no lysis or permeabilization steps are required. If cathepsin enzymes are active, the Magic Red substrate is cleaved and the cresyl violet fluorophore 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 with a fluorescence plate reader. Hoechst 33342 and Acridine Orange are included in the BioAssay™ kit to detect nuclear morphology and lysosomal organelle structure, respectively.||Kit Components:|25 tests:|Magic Red Substrate (MR-RR2), 1 25 Test Vial|Hoechst 33342, 1x1mL|Acridine Orange, 1x0.5mL||100 tests:|Magic Red Substrate (MR-RR2), 1 100 Test Vial|Hoechst 33342, 1x1mL|Acridine Orange, 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).