High Throughput Screen for Caspase 3/7 Detection – APO HTS

Key Benefits:
  • Homogenous assay for active caspase 3/7.
  • Breakthrough in cell lysis buffer and preservation of caspase activity.
  • Results in a no-wash, one-step assay.
  • No need to wash out media from cell samples, just add the reagent directly to your experimental samples.
  • Easy to Use: No need to make cell lysates or run Western blots.
  • Works with suspension and adherent cells.
Assay Principle

Cell Technology’s APO 3/7 HTS Assay utilizes the quenched (z-DEVD)2-R110 peptide substrate for caspase 3/7 detection. The absorption and emission properties of the R110 dye are suppressed when attached to the z-DEVD peptide sequence. When R110 is cleaved away, by active caspase3/7, form the quenching DEVD sequence, the free dye excites at 488nm and emits at 515-530 nm. As a result of a novel and proprietary Lysis Buffer System, the APO 3/7 HTS Assay is a homogenous platform that can be utilized for high throughput fluorescence plate reader applications. The reagent is directly added to the samples thus eliminating any wash steps.

Apo3HTS


In this figure, Jurkat cells were stimulated with various concentrations of staurosporine for 3 hours, after which caspase 3/7 activity was analyzed using the APO 3/7 HTS kit.


Citations

Identification of single-domain, Bax-specific intrabodies that confer resistance to mammalian cells against oxidative-stress-induced apoptosis - Deyzi Gueorguieva, Shenghua Li, Nicole Walsh, Amit Mukerji, Jamshid Tanha, and Siyaram Pandey - The FASEB Journal 2006

Effects of Pacing Site on Left Ventricular Activation Sequences Using a Non-Contact Mapping System: Implications for Heart Failure Pacing - D. Xing, F. Devecchi, T. Staley, D. Glassman, J. Martins

Role of Shear Stress in the differential regulation of endothelial Cathepsins and Cystatin C - PhD dissertation - Manu Omar Platt

Inhibitor of Metalloproteinases 3 Regulates Resolution of Inflammation following Acute Lung Injury- Sean E. Gill, Isham Huizar, Eli M. Bench, Samuel W. Sussman, Ying Wang, Rama Khokha, and William C. Parks - American Journal of Pathology 2010;176:64-73

References:
  1. 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.
  2. 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.
  3. 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.
  4. 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.
  5. Kumar, S. 1999. Mechanisms mediating caspase activation in cell death. Cell Death and Differ. 6: 1060-1066.
  6. Alnemri, E.S. et al (1996) Cell 87:171
  7. Trends Biochem Sci 22,388 (1997)
Kit contents
The APO HTS kit includes the following:
  • Part #4004: 1 vial caspase 3/7 reagent (z-DEVD)2 Rodamine 110
  • Part #3005 1 bottle of cell lysis buffer
Kit Availability
The following APO HTS kits are available:

 ProductCatalog #SizePrice
High Throughput Screen for Caspase 3/7 activityAPO200-2100$275.00
High Throughput Screen for Caspase 3/7 activityAPO200-3500$945.00
High Throughput Screen for Caspase 3/7 activityAPO200-41000$1,545.00

* Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for volume pricing