Flow Cytometry Immunofluorescence Staining Protocols

Protocol for flow cytometric determination of leukocyte surface antigens in whole blood. Includes: Blood collection; Add antibody to the blood; Make measurements. - [Read Flow Cytometric Determination of Leukocyte Surface Antigens in Whole Blood Protocol]

Protocol for fluorescein labeling of proteins. - [Read Fluorescein Labeling of Proteins Protocol]

Fluorescent dyes absorb light at certain wavelengths and in turn emit their fluorescence energy at a higher wavelength. Each dye has a distinct emission spectrum, which can be exploited for multicolor analysis. eBioscience antibodies are available conjugated to a wide variety of fluorochromes. - [Read Fluorescent Dyes for Flow Cytometric Analysis]

Protocol provides a method for analysis of cell surface antigens using direct immunofluorescence, i.e., when the antibody being used is directly labeled to a fluorescent dye. After staining, the cells can be analyzed immediately by flow cytometry or fixed and stored for later analysis. - [Read Fluorescent Staining of Cell Surface Antigens on Viable Cells Using Direct Immunofluorescence]

Protocol for four color immunophenotyping on the FACSCalibur. - [Read Four Color Immunophenotyping on the FACSCalibur Protocol]

Protocol for immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry. - [Read Immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry Protocol]

Minimizing background in immunofluorescence procedures. - [Read Minimizing Background in Immunofluorescence Procedures]

Provides a brief historical perspective to illustrate the demands for this technology and to lay the foundation for its application; Explains the hurdles that were surmounted to achieve the current level of multiparametric analysis which serves to alert researchers to potential problems they may encounter when they either bring this technology to their own laboratories, or when they use extant instrumentation in another laboratory; Illustrates some of the complexities that arise. - [Read Multiparameter FACS Analysis]

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]

Protocol for simultaneous measurement of cell surface immunofluorescence, viability, and DNA content. - [Read Simultaneous Measurement of Cell Surface Immunofluorescence, Viability, and DNA Content Protocol]

Protocol for simultaneous measurement of dual-color immunofluorescence and DNA/RNA content. - [Read Simultaneous Measurement of Dual-Color Immunofluorescence and DNA/RNA Content Protocol]

Protocol for simultaneous measurement of three-color immunofluorescence and DNA content. - [Read Simultaneous Measurement of Three-Color Immunofluorescence and DNA Content Protocol]