Different cell types vary in their phosphatidylserine (PS) content, along with the amount of PS exposure on the cell surface after cell death. This protocol is a guideline for getting started, however it may be necessary to adjust the concentration of the Annexin V-FITC. - [Read Annexin V Protocol for Flow Cytometry]
Protocol uses the BIOPRIME reaction kit from GibcoBRL to prepare biotin-labelled BAC DNA which is detected using FITC-Avidin (Vector Labs, DCS grade). Reagents from other manufacturers may work equally well but have not been tested. Includes: Labeling of BAC clones; Ethanol precipitation; Hybridization; Post-hybridisation treatment / detection. - [Read BAC-FISH Protocol]
Protocol uses specific antibodies coupled to one of four fluorochromes: fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), peridinin chlorophyll-a (PcP),
and allophycocyanin (APC). These fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The
fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]
Detection of Phosphatidylserine Externalization During Apoptosis. Shailaja Kasibhatla et al. An early event in apoptosis is the externalization of phosphatidylserine (PS), a phospholipid normally restricted to the inner leaflet of the plasma membrane. This apoptotic event can be monitored using Annexin V, a PS-specific binding protein. This protocol uses Annexin V-FITC as a probe, but Annexin V-biotin is also available, and binding can be revealed using streptavidin-FITC or oth - [Read Detection Of Phosphatidylserine Externalization During Apoptosis (Subscription Required)]
Protocol for phenotype-specific immunodetection of cyclins using 488/630 nm dual laser flow cytometry. This protocol is for use with the D and E cyclins and employs 488 nm argon laser excitation of propidium iodide and a FITC-conjugated phenotypic label, and 630 nm NeNe or diode laser excitation of the fluorochrome Cy5 to detect cell cycle-specific cyclin D expression. - [Read Phenotype-Specific Immunodetection of Cyclins using 488/630 nm Dual Laser Flow Cytometry Protocol]