Apoptosis Cytometry Protocols

Annexin V, belonging to a recently discovered family of proteins, the annexins, with anticoagulant properties has proven to be a useful tool in detecting apoptotic cells since it preferentially binds to negatively charged phospholipids like PS in the presence of Ca2+ and shows minimal binding to phosphatidylcholine and sphingomyeline. Changes in PS asymmetry, which is analyzed by measuring Annexin V binding to the cell membrane, were detected before morphological changes associated with... - [Read Annexin V Protocol]

Apoptosis detection using the protocol annexin V + 7-AAD. - [Read Apoptosis Detection: Annexin V + 7-AAD Protocol]

Protocol for apoptosis detection using Hoechst vs. 7-AAD protocol. - [Read Apoptosis Detection: Hoechst vs. 7-AAD Protocol]

Protocol for apoptosis detection using TUNEL protocol. This technique uses the enzyme terminal deoxynucleotidyl transferase (TdT) to label cells that have oligonucleosomal nicks/strand breaks in their DNA. - [Read Apoptosis Detection: TUNEL Protocol]

An integrative procedure through which a relatively satisfactory result can be obtained following a single stage of cell culture and transient cell treatment, then detection with different instruments. This shortens experiment time. - [Read Apoptosis Identification and Measurement Protocol]

Protocol for the detection of BrdU incorporation in DNA synthesizing cells. - [Read Detection of BrdU Incorporation in DNA Synthesizing Cells Protocol]

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. - [Read Flow Cytometry of Apoptosis Protocol]

Protocol for measuring apoptosis and necrosis by dual laser flow cytometry. Includes: Preparation of stock solutions of HO342, PI, and 7-AAD; Staining for detection of apoptosis; - [Read Measuring Apoptosis and Necrosis by Dual Laser Flow Cytometry Protocol]

Propidium iodide (PI) intercalates into double-stranded nucleic acids. It is excluded by viable cells but can penetrate cell membranes of dying or dead cells. - [Read Propidium Iodide Staining of Dead Cells for Flow Cytometry Protocol]

The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and the Cell Cycle Distribution of Primary B Cells Using PI]

Two distinct modes of cell death, apoptosis and necrosis, can be distinguished on the basis of differences in morphological, biochemical, and molecular changes occurring in the dying cells. Report the development of flow cytometric techniques that permit concomitant detection of apoptosis and cellular DNA content or BrdU content analysis. - [Read The Use of Flow Cytometry for Concomitant Detection of Apoptosis and Cell Cycle Analysis]