Real-time detection of cellular apoptosis using a rat C6 glioma cell-based assay system

Manager 2019.03.07 14:00 Views : 770

Year 2011 
Journal Info. Molecular & Cellular Toxicology, 7, 177-184 (2011). 
Author Kyoung Hwa Jung*, Young Me Song*, Nando Dulal Das, Kyoung Sun Park, Mi Ran Choi, Sang Youn Hwang, Eun Kyu Lee, Moon Kwon Lee, Jaebum Choo, Kyoung Suk Kim, Moo Soung Kim, Sang Rin Lee, Young Gyu Chai 
etc.  

Abstract

Caspase-3 is a key mediator of apoptosis in mammalian cells. Cells expressing caspase-3 substrate peptides have become powerful and increasingly common components of cell-based assay systems. We developed a cell-based assay to measure staurosporine (STP)-induced caspase-3 activity in live rat glioma C6 cells. The caspase-3 sensing system was constructed to include a nuclear export signal (NES), followed by the amino acid sequence Asp-Glu-Val-Asp (DEVD) and a green fluorescent protein (GFP) fused to the Nterminal site of a nuclear localization signal (NLS). Using time-lapse confocal microscopy, we monitored caspase-3 activation during apoptosis by imaging the translocation of GFP from the cytosol to the nucleus. After 8 h of 0.5 μM STP treatment, caspase-3 activity was assessed by monitoring the translocation of GFP to the nucleus due to cleavage of the NES from the GFP by caspase-3. Finally, disintegration of the plasma membrane during late apoptosis was confirmed using a nuclear dye, propidium iodide. Analysis of caspase-3 activity using real-time monitoring can potentially be used to screen for apoptotic molecules in living cells.

 

Download PDF = Click Attachment, please.

I want to
List
Year Title Journal Info. etc.
2011  Real-time detection of cellular apoptosis using a rat C6 glioma cell-based assay system file Molecular & Cellular Toxicology, 7, 177-184 (2011).   
2011  Ultraviolet Absorption Spectra, Structure, Vibrations, and Theoretical Calculations of 2-Fluoro- and 3-Fluoropyridine in their Electronic Excited States file Chemical Physics Letters. 514, 214-219 (2011).   
2011  Polycrystalline tungsten oxide nanofibers for gas-sensing application file Sensors and Actuators B, 160, 549-554 (2011).   
2011  Simultaneous immunoassay for the detection of two lung cancer markers using functionalized SERS nanoprobes file Chemical Communications. 47, 12515-12517 (2011).   
2011  Recent advances in microfluidic technologies for biochemistry and molecular biology file BMB reports, 44, 705-712 (2011).  review paper 
2011  Highly reproducible immunoassay of cancer markers on a gold-patterned microarray chip using surface-enhanced Raman scattering imaging file Biosensors and Bioelectronics, 26, 2135–2141 (2011).   
2011  Polymorph-tuned Synthesis of a- and b-Bi2O3 Nanowires and Determination of Their Growth Direction from Polarized Raman Single Nanowire Microscopy file Chemistry - A European Journal, 17, 1304-1309 (2011).   
2011  Au Nanowire-on-Film SERRS Sensor for Ultrasensitive Hg2+ Detection file Chemistry - A European Journal, 17, 2211-2214 (2011).   
2011  Pep-1 Peptide-Modified Liposomal Carriers for Intracellular Delivery of Gold Nanoparticles file Chemical & Pharmaceutical Bulletin, 59, 109-112 (2011).   
2011  Pt nanoparticle-supported multiwall carbon nanotube electrodes for amperometric hydrogen detection file Sensors and Actuators B, 155, 191-198 (2011).   
2011  Enzyme kinetic measurements using a droplet-based microfluidic system with a concentration gradient file Analytical Chemistry, 83, 1603-1608 (2011).   
2010  Fabrication of protein-anchoring surface by modification of SiO2 with liposomal bilayer file Colloids and Surfaces B: Biointerfaces, 75, 209-213 (2010).   
2010  Optofluidic platforms based on surface-enhanced Raman scattering file Analyst, 135, 837–844 (2010).  Cover page 
2010  Preparation of Targeting Proteoliposome by Postinsertion of a Linker Molecule Conjugated with Recombinant Human Epidermal Growth Factor file Bioconjugate Chemistry, 21, 345-351 (2010).   
2010  Nanomaterial-assisted aptamers for optical sensing file Biosensors and Bioelectronics, 25, 1859–1868 (2010).   
2010  High sensitive detection of C-reactive protein by total internal reflection fluorescence microscopy on rapidly making nanoarray protein chip file Talanta, 81, 1402–1408 (2010)   
2010  Effect of cholesterol content on affinity and stability of factor VIII and annexin V binding to a liposomal bilayer membrane file Chemistry and Physics of Lipids, 163, 335-340 (2010).   
2010  Electrochemical Patterning of Transparent Single-Walled Carbon Nanotube Films on Plastic Substrates file Langmuir, 26, 9136-9141 (2010).   
2010  On-chip Immunoassay Using Surface-Enhanced Raman Scattering of Hollow Gold Nanospheres file Analytical Chemistry, 82, 5290–5295 (2010).   
2010  Quantification of UV-induced cyclobutane pyrimidine dimers using an oligonucleotide chip assay file Analytical and Bioanalytical Chemistry, 397, 2271-2277 (2010).   
Write Tag