- Additional information
- ASSAY PRINCIPLE
- Readable Documents
- Kit contents and storage
- Cell permeability allow direct measurement of apoptosis and mitochondrial potential in live cells.
- Applications – Cells can be analyzed by Flow Cytometry, Fluorescent plate reader or Fluorescent microscopy.
- Incubate for 15 minutes, wash and measure.
- Add this reagent directly to live cells in your media of choice.
Detection of the mitochondrial permeability transition event provides an early indication of the initiation of cellular apoptosis. This process is typically defined as a collapse in the electrochemical gradient across the mitochondrial membrane, as measured by the change in the membrane potential (YD). Loss of mitochondrial (YD) is indicative of apoptosis and can be detected by a unique fluorescent cationic dye, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl- benzamidazolocarbocyanin iodide, commonly known as JC-1. This dye has been incorporated into the user-friendly kit for the simple and reproducible detection of the membrane potential (YD) event in apoptotic cells. The kit has been formatted for use on Flow cytometers, Fluorescent plate readers and Fluorescent Microscopes
Fig (A). Jurkat cells were cultured with DMSO for 2 hours. Cells were then stained with JC-1 Mitochondrial Membrane Potential Detection Kit for 15 minutes and analyzed by flow cytometry.
Fig (B). urkat cells were cultured with staurosporine for 2 hours. Cell were then stained with JC-1 Mitochondrial Membrane Potential Detection Kit for 15 minutes and analyzed by flow cytometry.
Kit contents and storage
|Part# 4001||Lyophilized JC-1 Dye||Store at 2-8°C|
|Part# 3002||10X Assay Buffer||Store at 2-8°C|
|Desagher, S., Osen-Sand, A., Nichols, A., Eskes, R., Montessuit, S., Lauper, S., Maundrell, K., Antonsson, B., and Martinou, J.C. Bid-induced conformational change of Bax is responsible for mitochondrial cytochrome c release during apoptosis. J. Cell Biol. 144 (5): 891-901 (1999).|
|Narita, M., Shimizu, S., Ito, T., Chittenden, T., Lutz, R. J., Matsuda, H., and Tsujimoto, Y. Bax interacts with the permeability transition pore to induce permeability transition and cytochrome c release in isolated mitochondria. Proc. Natl. Acad. Sci. USA 95: 14681-14686 (1998).|
|Basanez, G., Nechushtan, A., Drozhinin, O., Chanturiya, A., Choe, E., Tutt, S., Wood, K. A., Hsu, Y. T., Zimmerberg, J., and Youle, R. J. Bax , but not Bcl-XL decreases the lifetime of planar phospholipid bilayer membranes at subnanomolar concentrations. Proc. Natl. Acad. Sci. USA 96: 5492-5497 (1999).|
|Luo, X., Budihardio, I., Zou, H., Slaughter, C., and Wang, X. Bid, a Bcl-2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 94: 481-490 (1998).|
|Smiley, S. T., Reers, M., Mottola-Hartshorn, C., Lin, M., Chen, A., Smith, T. W., Steele, G.D., and Chen, L. B. Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate forming lipophilic cation JC-1. Proc. Natl. Acad. Sci. USA 88: 3671-3675 (1991).|
|Cossarizza, A., Baccarani-Contri, M., Kalashnikova, G., and Franceschi, C. A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5’,6,6’-tetrachloro-1,1’,3,3’-tetraethylbenzimidazolylcarbocyanine iodide (JC-1). Biochem. Biophys. Res. Commun. 197 (1): 40-45 (1993).|
|Reers, M., Smith, T. W., and Chen, L. B. J-aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential. Biochemistry 30: 4480-4486 (1991).|
|White, R. J., and Reynolds, I. J. Mitochondrial depolarization in glutamatestimulated neurons: an early signal specific to excitotoxin exposure. Journal of Neuroscience 16: 5688-5697 (1996).|
|Defects in Cell Growth Regulation by C 18:0-Ceramide and Longevity Assurance Gene 1 (LAG1) in Human Head and Neck Squamous Cell Carcinomas (HNSCC)||Serap Koybasi, Can E. Senkal, Kamala Sundararaj, et al||JBC Papers in Press||Published on August 17, 2004 as Manuscript M406920200|
|Effects of a series of organosulfur compounds on mitotic arrest and induction of apoptosis in colon cancer cells||http://mct.aacrjournals.org/cgi/content/full/4/9/1388||Danhua Xiao, John T. Pinto, Gregg G. Gundersen & Bernard Weinstein||Mol Cancer Ther||2005;4:1388-1398|
|Adrenergic receptor-stimulated apoptosis in adult cardiac myocytes involves MMP-2-mediated disruption of β1 integrin signaling and mitochondrial pathway||http://ajpcell.physiology.org/cgi/content/abstract/290/1/C254||Bindu Menon,Mahipal Singh, Robert Ross, Jennifer N. Johnson and Krishna Singh||Am J Physiol Cell Physiol||290: C254-C261, 2006. First published September 7, 2005|
|SGLT-1-mediated glucose uptake protects intestinal epithelial cells against LPS-induced apoptosis and barrier defects: a novel cellular rescue mechanism?||http://www.fasebj.org/cgi/content/abstract/19/13/1822||Linda C. H. Yu Andrew N. Flynn, Jerrold R. Turner and Andre G. Buret||The FASEB Journal||2005;19:1822-1835|
|Ceramide induces mitochondrial abnormalities in insulin-secreting INS-1 cells: Potential mechanisms underlying ceramide-mediated metabolic dysfunction of the β cell||http://www.springerlink.com/content/t3g0p313518r2747/||R. Veluthakal, R. Palanivel Y. Zhao, P. McDonald, S. Gruber and A. Kowluru||Apoptosis Journal||Vol 10/No 4, Aug 2005|
|Opposing effects of bovine papillomavirus type 1 E6 and E7 genes on Fas-mediated apoptosis||http://www.nature.com/onc/journal/v24/n24/abs/1208542a.html||Yun Liu, Zhiguo Liu, Hua Gao, You Zhou, Elliot J Androphy and Jason J Chen||Oncogene||(March 2005) 24, 3942–3953|
|Resveratrol-caused apoptosis of human prostate carcinoma LNCaP cells is mediated via modulation of phosphatidylinositol 3′-kinase/Akt pathway and Bcl-2 family proteins||http://mct.aacrjournals.org/cgi/content/abstract/5/5/1335||Moammir H. Aziz,Minakshi Nihal, Vivian X. Fu, David F.Jarrard and N Ahmad||Mol Cancer Ther||2006;5:1335-1341|