Apo Active 3 FITC

Product Description

Key Benefits

• Highly specific for active human and murine caspase 3. Other assays require the utilization of peptide based (DEVD) reagents that tend to cross-react with caspase 7 and other caspases.
• Applications – Works with a fluoresence microscope, 96 well plate reader, or flow cytometer.
• Yields both quantitative and qualitative results. Gives a strong positive signal.
• Can be used in conjunction with other antibodies or stains.
• No need to make cell lysates or run western blots. Cells can be fixed and analyzed later.
• Works with human, mouse and rat cell lines.

Assay Principle

Cell Technology ’s APO-ACTIVE CASPASE 3 Kit utilizes a rabbit affinity purified polyclonal antibody raised against amino acid 163-175 of murine caspase 3 (9). This neo epitope is present on the p18 subunit of cleaved caspase 3 (9). Cells undergoing apoptosis are fixed and permeablized prior to the addition of the antibody. A secondary FITC labeled goat anti rabbit antibody is used to visualize the bound rabbit anti caspase 3 polyclonal.

Fig (A). Jurkat Cells were stimulated with DMSO for 2 hours. The cells were washed and fixed for 15 minutes. The cells were then permeabilized with saponin and stained with Rabbit anti active caspase 3 antibody for 1 hour. The samples were then stained with PE labeled Goat anti Rabbit antibody.

Fig (B). Jurkat Cells were stimulated with staurosporine for 2 hours. The cells were washed and fixed for 15 minutes. The cells were then permeabilized with saponin and stained with Rabbit anti active caspase 3 antibody for 1 hour. The samples were then stained with FITC labeled Goat anti Rabbit antibody.

Citations

The endoplasmic reticulum is the site of cholestrol-induced cytotoxicity in macrophages - Feng, Yao, Li, Devlin et al - Nature Cell Biology Vol 5, No 9 September 2003, pp 781-792

Combination chemotherapy with gemcitabine and biotherapy with opioid growth factor (OGF) enhances the growth inhibition of pancreatic adenocarcinoma - Ian S. Zagon, Jeffrey R. Jaglowski, et.al. - - Cancer Chemotherapy and Pharmacology Vol 56/ No 5, Nov 2005

The Pseudomonas aeruginosa type III secreted toxin ExoT is necessary and sufficient to induce apoptosis in epithelial cells - Sasha H. Shafikhani, Christina Morales and Joanne Engel - Cellular Microbiology Vol 10 Issue 4, pp 994-1007, Dec 2007

Monoclonal Anti–Interleukin 23 Reverses Active Colitis in a T Cell–Mediated Model in Mice - C. Elson, Y. Cong, C. Weaver, T. Schoeb, T. McClanahan, R. Fick, R. Kastelein - Gastroenterology Volume 132, Issue 7, Pages 2359-2370 (2007)

Acute and chronic microvascular alterations in a mouse model of ischemic acute kidney injury - Markus Horbelt, So-Young Lee, Henry E Mang, Nicole L. Knipe, Yoshikazu Sado, Andreas Kribben, and Timothy A Sutton - Am J Physiol Renal Physiol July 11, 2007

Cytotoxicity and Secretion of Gamma Interferon Are Carried Out by Distinct CD8 T Cells during Mycobacterium tuberculosis Infection - Thorbjorg Einarsdottir, Euan Lockhart, and JoAnne L. Flynn - American Society for Microbiology - Infection and Immunity October 2009, p. 4621-4630, Vol. 77, No. 10.

Converting cell lines representing hematological malignancies from glucocorticoid-resistant to glucocorticoid-sensitive: Signaling pathway interactions - Anna S. Garza, Aaron L. Miller, Betty H. Johnson, E. Brad Thompson - Leukemia Research Vol Vol 33, Issue 5, pp 717-727. May 2009

References

• 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.
• Alnemri, E.S. et al (1996) Cell 87:171
• Trends Biochem Sci 22,388 (1997)
• Nature 376, 37 (1995).
• Srinivasan A, et al Cell Death and Differentiation (1998) 5: 1004-1016

Kit contents and Long Term storage