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Technical guide to Cell Proliferation and Apoptosis Methods (Roche) hot top rated recommended protocol
This is a technical guide to Cell Proliferation and Apoptosis Methods (Roche). Guide to Cell Proliferation and Apoptosis Methods Table of Contents Chapter 1: Cell Death - Apoptosis and Necrosis 1.1 Introduction 2 1.1.1 Terminology ...

In Situ Cell Death (Apoptosis) Detection by TUNEL labeling new protocol
In Situ Cell Death (Apoptosis) Detection by TUNEL labeling protocol. A modified Boehringer Mannheim (Catalog No. 1684809) protocol by Josiah N. Wilcox and Jos¨¦ C. Rodriguez - Emory University. ...

In Situ Cell Death (Apoptosis) Detection by TUNEL labeling on Paraffin section protocol
A modified Boehringer Mannheim (Catalog No. 1684809) protocolby Josiah N. Wilcox, Jos¨¦ C. Rodriguez, and H¨¦ctor de L¨¦on - Emory University. ...

General Annexin V Staining Procedure protocol
General Annexin V Staining Procedure from BD Pharmingen. ...

Radioactive DNA Fragmentation Assay protocol
The DNA Fragmentation Assay allows to determine the amount of DNA that is degraded upon treatment of cells with certain agents, e.g. with TNF-alpha or anti-Fas antibody (IPO-4). The protocol gives description, principle of the assay and procedure of DNA fragme ...

DNA laddering assay for treated cells protocol
DNA laddering can be detected from samples with only 8% apoptotic cells. Alternatively, the cells can be stained with DAPI and analysed by flow cytometry. (CellDeath.de) ...

Gentle lysis of mammalian cells for cytochrome c release assay protocol
Lysis in the sucrose-containing Buffer ("Mito-Buffer") is supposed to prevent accidential disrupture of the mitochondria to prevent the leakage of mitochondrial proteins (such as cytochrome c) into the cytosol. (CellDeath.de) ...

Caspase-3 Fluorometric Assay new protocol
This protocol describes detailed procedures on determination of the increased enzymatic activity of the caspase-3 class of proteases in apoptotic cells by fluorometric reaction. (R&D systems) ...

Commonly used Cytometry Apoptosis Detection Techniques new protocol
Hoechst and 7-AAD H342 is DNA stain that binds preferentially to A-T base-pairs without the necessary of permeabilization step. However, it is necessary to set up incubation conditions for efficient internalization of the dye. Hoechst 33342 (Bisbenzi ...

apoptosis resource collections site
This page contains information about apoptosis overview, developmental cell death, apoptosis pathway, apoptosis protocols, apoptosis journal & literature database. (IHC world) ...

Apoptosis Assistant (Promega) software
Apoptosis Assistant (Promega) Apoptosis Assistant helps you find experimental information from the literature that has used our products to detect apoptosis in cell lines and tissues. This tool contains information on cell lines, the type of reagent ...

Apoptosis pathway images and posters new media
Here is the collection of apoptosis pathway illustrations. Some of them are available in poster when requested. Most of pathway illustrations give general overview of the apoptosis pathways. ...

More apoptosis assay protocols

Apoptosis protocols
(Curr Protoc Cell Biol.)
1: Curr Protoc Cell Biol. 2004 Feb;Chapter 18:Unit 18.8.

Flow cytometry of apoptosis.

Pozarowski P, Grabarek J, Darzynkiewicz Z.

School of Medicine, Lublin, Poland.

Common methods applicable to flow cytometry make it possible to: (1) identify and
quantify dead or dying cells, (2) reveal a mode of cell death (apoptosis or
necrosis), and (3) study mechanisms involved in cell death. Gross changes in cell
morphology and chromatin condensation, which occur during apoptosis, can be
detected by analysis with laser light beam scattering. Early events of apoptosis,
dissipation of the mitochondrial transmembrane potential and caspase activation,
can be detected using either fluorochrome reporter groups or appropriate
antibodies. Exposure of phosphatidylserine on the exterior surface of the plasma
membrane can be detected by the binding of fluoresceinated annexin V. Another
apoptotic event, DNA fragmentation based on DNA content of cells with fractional
("sub-G1") or DNA strand-break labeling, TUNEL; or In Situ End Labeling, ISEL;.
Still another hallmark of apoptosis is the activation of tissue transglutaminase
(TGase), the enzyme that crosslinks protein and thereby makes them less
immunogenic. The major advantage of flow cytometry in these applications is that
it provides the possibility of multiparametric measurements of cell attributes.

2: Curr Protoc Cell Biol. 2001 Nov;Chapter 18:Unit 18.3.

Assessment of apoptosis and necrosis by DNA fragmentation and morphological
criteria.


Zhivotosky B, Orrenius S.

Karolinska Institute, Stockholm, Sweden.

Apoptotic cells share a number of common features, such as phosphatidylserine
(PS) exposure, cell shrinkage, chromatin cleavage, nuclear condensation, and
formation of pyknotic bodies of condensed chromatin. Necrotic cells exhibit
nuclear swelling, chromatin flocculation, loss of nuclear basophilia, breakdown
of cytoplasmic structure and organelle function, and cytolysis by swelling. This
unit describes some of the techniques most commonly used to detect cell death. A
number of assays are used for characterizing and distinguishing apoptosis and
necrosis. Morphological assays include trypan blue exclusion, differential
staining, and Hoechst staining. Methods to detect chromatin cleavage include
TUNEL assays for whole cells and paraffin sections, DNA fragmentation assays
using whole cells, assays of total genomic DNA, analysis of DNA fragmentation by
agarose gel electrophoresis, phenol extraction of DNA for analysis of
fragmentation, a quantitative assay for DNA fragmentation, and detection of DNA
fragmentation by pulsed-field gel electrophoresis. A protocol is also provided
for Cytospin preparations from cell suspensions.

3: Curr Protoc Cell Biol. 2001 Aug;Chapter 18:Unit 18.2.

Analysis of caspase activation during apoptosis.

Kaufmann SH, Kottke TJ, Martins LM, Henzing AJ, Earnshaw WC.

Mayo Clinic, Rochester, Minnesota, USA.

This unit describes three methods for the detection of caspase activation as
cells undergo apoptosis. Simple and relatively quantitative enzymatic assays are
provided using suitable substrates. Because the various low-molecular-weight
substrates available for these assays are not selective, however, the assays do
not accurately distinguish between various caspases. Immunoblotting is described
for following the activation of specific caspases. When coupled with subcellular
fractionation, this method can provide large amounts of temporal and spatial
information about caspase activation. Finally, affinity labeling protocols are
provided for detecting active caspases in whole-cell lysates or subcellular
fractions.

4: Curr Protoc Cell Biol. 2001 Aug;Chapter 18:Unit 18.1.

Current concepts in cell death.

Zhivotovsky B, Orrenius S.

Karolinska Institutet, Stockholm, Sweden.

This unit attempts to define cell toxicity that leads to cellular demise, with a
strong emphasis on cell death via both apoptosis and necrosis, by summarizing
some of the more recent developments in cellular, molecular, and biochemical
studies of the events that govern the induction and execution of cell death.
Specific topics pertaining to cell death include structural changes,
macromolecular degradation, cellular signaling, the role of mitochondria, and
genetic modulation of apoptotic cell death.

Cell cycle books
(Biowww Bookshelf)
Cell cycle related research

Apoptosis and cell proliferation books
(Biowww bookshelf)
apoptosis and proliferation research books

Figure. Fragmented DNA can be labeled by terminal deoxynucleotidyl transferase (TdT) for apoptosis
(Immunobiology)
Appendix I. Immunologists' Toolbox Characterization of lymphocyte specificity, frequency, and function.

Figure. The caspase cascade involved in apoptosis
(Molecular Biology of the Cell)
IV. Internal Organization of the Cell 17. The Cell Cycle and Programmed Cell Death Programmed Cell Death (Apoptosis)

Programmed Cell Death (Apoptosis)
(Molecular Biology of the Cell)
IV. Internal Organization of the Cell 17. The Cell Cycle and Programmed Cell Death

Death Domain Proteins
(Cancer Medicine)
Part II Scientific Foundation, Section 1: Cancer Biology 4. Apoptosis and Cancer

BrdU/TUNEL double staining
(Forum)
endothelial cells double staining for BrdU and TUNEL


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