产品资料

Cell Technology/ApoLogix SR-FMK/25/SR100

Description
Additional Information
Readable Documents
Assay Principle
Reviews

Key Benefits

Non-cytotoxic assay arrests further apoptotic activity via caspase inhibition.
Cell permeablity permits direct visualization of cytosolic apoptotic events.
Apoptotic cell population does not diminish over time.
Add reagent directly to cells. No special buffer or media needed. No preparation of cell lysates required. Simple wash procedure.
Works in diverse cell lines: human, rodent, Drosophila.
Can be performed in conjunction with Annexin staining, TUNEL, antibody staining, or with other APO LOGIX reagents on the same population of cells.
Permits high through-put screening. Protocol can be adapted for ex vivo as well as in situ experiments.
Applications – Works with fluorescence microscope, 96-well fluorescence plate readers
Yields both quantitative and qualitative results. Gives strong signal with little background noise.

Additional information

Kit Size	25, 100

APO LOGIX SR kits contain a generic sulforhodamine labeled caspase inhibitor (sulforhodamine-peptide-fluoromethyl ketone). This reagent is cell permeable and is used on whole cells to detect apoptosis. Apoptotic cells are detected by a fluorescence plate reader or fluorescence microscope using an excitation source at 550nm and measuring emission at 595nm. The assay takes about 1 hr to completeAPO LOGIX Sulforhodamine

Jurkat cells stimulated with staurosporine for 2 hours and then labeled with SR-VAD-FMK.

Left side: 30X phase contrast

Right side: 30X fluorescence microscope. Excitation: 550nm emission > 580nm.APO LOGIX Sulforhodamine

Jurkat cells stimulated with staurosporine for 2 hours. Cells were then stained with SR-VAD-FMK for 1 hour and read in a 96 well fluorescence plate reader.

Document Title

SR protocol

SRVADFMK Datasheet

msds.Apologix

Reference

Slee, E. A., C. Adrain, and S. J. Martin. 1999. Serial Killers: ordering caspase activation events in apoptosis. Cell Death and Differ. 6:1067-1074.

Walker, N. P., R. V. Talanian, K. D. Brady, L. C. Dang, N. J. Bump, C. R. Ferenz, S. Franklin, T. Ghayur, M. C. Hackett and L. D. Hammill. 1994. Crystal Structure of the Cysteine Protease Interleukin-1ß-Converting Enzyme: A (p20/p10)2 Homodimer. Cell 78:343-352.

Wilson, K. P., J. F. Black, J. A. Thomson, E. E. Kim, J. P. Griffith, M. A. Navia, M. A. Murcko, S. P. Chambers, R. A. Aldape, S. A. Raybuck, and D. J. Livingston. 1994. Structure and mechanism of interleukin-1 beta converting enzyme. Nature 370: 270-275.

Rotonda, J., D. W. Nicholson, K. M. Fazil, M. Gallant, Y. Gareau, M. Labelle, E. P. Peterson, D. M. Rasper, R. Ruel, J. P. Vaillancourt, N. A. Thornberry and J. W. Becker. 1996. The three-dimensional structure of apopain/CPP32, a key mediator of apoptosis. Nature Struct. Biol. 3(7): 619-625.

Kumar, S. 1999. Mechanisms mediating caspase activation in cell death. Cell Death and Differ. 6: 1060-1066.

Thornberry, N. A., T. A. Rano, E. P. Peterson, D. M. Rasper, T. Timkey, M. Garcia-Calvo, V. M. Houtszager, P. A. Nordstrom, S. Roy, J. P. Vaillancourt, K. T. Chapman and D. W. Nicholson. 1997. A combinatorial approach defines specificities of members of the caspase SRily and granzyme B. Functional relationships established for key mediators of apoptosis. J. Biol. Chem. 272(29): 17907-17911.

Amstad, P.A., G.L. Johnson, B.W. Lee and S. Dhawan. 2000. An in situ marker for the detection of activated caspases. Biotechnology Laboratory 18: 52-56.

Bedner, E., P. Smolewski, P.A. Amstad and Z. Darzynkiewicz. 2000. Activation of caspases measured in situ by binding or fluorochrome-labeled inhibitors of caspases (FLICA): correlation with DNA fragmentation. Exp. Cell Research 259: 308-313.

Smolewski, P., E. Bedner, L. Du, T.-C. Hsieh, J. Wu, J. D. Phelps and Z. Darzynkiewicz. 2001. Detection of caspase activation by fluorochrome-labeled inhibitors: multiparameter analysis by laser scanning cytometry. Cytometry 44: 73-82.

Ekert, P. G., J. Silke and D. L. Vaux. 1999. Caspase inhibitors. Cell Death and Differ. 6:1081-1086.

Carcia-Calvo, M., E. Peterson, B. Leiting, R. Ruel, D. Nicholson and N. Thornberry. 1998. Inhibition of human caspases by peptide-based and macromolecular inhibitors. J. Biol. Chem. 273: 32608-32613.

Hirata, H., A. Takahashi, S. Kobayashi, S. Yonehara, H. Sawai, T. Okazaki, K. Yamamoto and M. Sasada. 1998. Caspases are activated in a branched protease cascade and control distinct downstream processes in Fas-induced apoptosis. J. Exp. Med. 187: 587-600

Part#	Reagent	Temperature
Part # 679	Lyophilized SR-VAD-FMK	2-8C
Part # 635	10X Wash Buffer	2-8C
Part # 636	10X Fixitive	2-8C

Please select an ACF field to output

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Cell Technology/ApoLogix SR-FMK/25/SR100

Key Benefits

Additional information