Summary: Cell death can occur by either of two distinct mechanisms, necrosis or programmed cell death (apoptosis). Necrosis is a pathological process which occurs when cells are exposed to a serious physical or chemical insult. Apoptosis is a physiological and controlled process by which unwanted or useless cells are eliminated during development and other normal biological processes. Apoptosis can be detected in populations of cells or in individual cells. Many different methods have been devised to detect apoptosis such as The TUNEL (TdT-mediated dUTP Nick-End Labeling) analysis, ISEL (in situ end labeling), and DNA laddering analysis for the detection of fragmentation of DNA in populations of cells or in individual cells, Annexin-V analysis that measures alterations in plasma membranes, detection of apoptosis related proteins such p53 and Fas.
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Protocols
- Apoptosis: A Laboratory Manual of Experimental Methods (Andrea Cossarizza & Diana Boraschi)
This online book covers essentially all experimental aspects of apoptosis.
http://imoax1.unimo.it/~cossarizza/apobook98/index...
Added: Fri Mar 26 2004, Hits: 1151, Reviews: 0 Write review Cached - Guide to Cell Proliferation and Apoptosis Methods
(Roche Diagnostics Corporation)
Comprehensive guide to cell prolifieration, cytotoxicity, apoptosis, including principles and methods. Major topics are Apoptosis Assay Methods/Cytotoxicity Assay Methods/Cell Proliferation and Viability.
http://www.roche-applied-science.com/sis/apoptosis...
Added: Fri Sep 20 2002, Hits: 9138, Reviews: 0 Write review Cached - Staining Methods for Cell Death (Steven Finkbeiner, UCSF)
Different staining methods to detect dead and apoptotic cells including trypan blue, FDA (fluorescein diacetate)-green, alive cells; P.I. (propidium iodide)-red, dead cells, Hoescht staining
http://www.gladstone.ucsf.edu/gladstone/files/fink...
Added: Fri Feb 20 2009, Hits: 7904, Reviews: 0 Write review -
A Rapid & Quantitative Method for Detecting Apoptosis by FACS in Transiently Transfected Cells
(Nucl. Acids Res)
a method suitable for the flow cytometric determination of apoptosis in a large number of samples by propidium iodide (PI) in a population of cells transiently co-transfected with a plasmid encoding enhanced green fluorescent protein (eGFP) and a plasmid carrying a gene of interest. This assay combines the FACS-optimised GFP mutant (eGFP) as a marker to detect transiently transfected cells and the reduced fluorescence of the DNA-binding dye PI in the apoptotic subpopulation as a marker for apoptosis.
http://nar.oxfordjournals.org/cgi/content/full/25/...
Added: Fri Mar 20 2009, Hits: 2726, Reviews: 0 Write review -
Apoptosis Miniassay
(Hannun/Obeid Lab)
http://www.musc.edu/bcmb/ceramide/protocols/0009.h...
Added: Tue May 14 2002, Hits: 3059, Reviews: 0 Write review Cached -
Cell death detection in Xenopus embryos by ELISA
(The Xenopus Molecular Marker Resource)
http://www.xenbase.org/xenbase/original/WWW/Marker...
Added: Sat Feb 21 2009, Hits: 1471, Reviews: 0 Write review Cached -
Detection of Apoptosis in Paraffin-embedded tissues by ISEL (In Situ End Labeling)
(LEP)
The ISEL technique identifies morphologically apparent karyorrhetic cells and also labels cells in which the DNA fragmentation has not yet progressed to frank nuclear fragmentation. By using biotinylated nucleotides and a DNA polymerase, the DNA fragments serve as templates in situ to synthesize new DNA strands, which are visualized by light microscopy after histochemical processing.
http://dir.niehs.nih.gov/dirlep/ish/isel1.html
Added: Tue May 14 2002, Hits: 2010, Reviews: 0 Write review Cached -
Induced Apoptosis Using Anti-Fas mAb
(eBioscience)
This is a simple yet powerful protocol to induce CD95 (Fas)-mediated apoptosis of human Jurkat cell line by monoclonal antibody EOS9.1. This can be used as a general positive control for apoptosis detection systems. For best results, cells should be in their exponential log phase with viability rate of greater than 95% when they are used for this assay.
http://www.ebioscience.com/ebioscience/appls/Apop....
Added: Tue May 14 2002, Hits: 1362, Reviews: 0 Write review Cached -
Induction of Apoptosis
(Oncogene)
General protocol for 48h DNA Damage-Induced Apoptosis. This protocol is based on p53-dependent G1-arrest that occurs in response to DNA damage by chemical agents such as doxorubicin, 5-fluorouracil, paclitaxel or vinblastine. A typical time course for p53 and p21WAF1 induction is 40 to 48 hours treatment with a DNA damaging agent.
http://www.oncresprod.com/techresources/TechTips.a...
Added: Tue May 14 2002, Hits: 2276, Reviews: 0 Write review Cached -
JC-1 Stain of Apoptotic Cells
(Research Group of Johannes Schmid, Medical University of Vienna)
JC1-staining to detect apoptotic cells via the mitochondrial membrane potential.
http://www.meduniwien.ac.at/user/johannes.schmid/J...
Added: Fri Nov 06 2009, Hits: 1925, Reviews: 0 Write review Cached -
Methods for Cell Death Staining
(Steven Finkbeiner, Departments of Neurology and Physiology, UCSF)
Includes trypan blue staining, propidium iodide staining of living cells and fixed cells, Hoescht staining.
http://gweb1.ucsf.edu/labs/finkbeiner/Protocol/pro...
Added: Tue Oct 01 2002, Hits: 1747, Reviews: 0 Write review Cached