Apo Active 3 FITC

$155.00$445.00

Clear

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.

Additional information

Kit Size

25, 100

ASSAY PRINCIPLE

apo-active-3-img1_

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.

apo-active-3-img2_

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.

Readable Documents

Title Name
Protocol Protocol.pdf
Datasheet Datasheet.pdf
msds msds.pdf

Kit contents and storage

Part# Reagent Temperature
Part # 1002 Rabbit Anti Cleaved Caspase 3 Affinity Purified Polyclonal Antibody, 1 Vial 2-8°C
Part # 2002 FITC Labeled Goat Anti Rabbit Affinity Purified Polyclonal Antibody, 1 Vial 2-8°C
Part # 3001 10X Fixative, 1 Vial 2-8°C

 

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

Citations

Title File Link Author(s) Journal Year; Edition:Pages
The endoplasmic reticulum is the site of cholestrol-induced cytotoxicity in macrophages http://www.cumc.columbia.edu/dept/medicine/tabas_site/pubs/UPR-NatureCB.pdf 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 http://www.springerlink.com/content/h025524175r45553/ 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 http://www3.interscience.wiley.com/journal/119420124/abstract?CRETRY=1&SRETRY=0 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 http://linkinghub.elsevier.com/retrieve/pii/S0016508507006300 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 http://ajprenal.physiology.org/cgi/content/abstract/00452.2006v1 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 http://iai.asm.org/cgi/content/abstract/77/10/4621 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 http://www.lrjournal.com/article/S0145-2126%2808%2900462-1/abstract Anna S. Garza, Aaron L. Miller, Betty H. Johnson, E. Brad Thompson Leukemia Research Vol Vol 33, Issue 5, pp 717-727. May 2009