Flow Cytometry Protocols
Staining cells with a No-Lyse protocol
Explore the step-by-step process for staining mononuclear cells using fluorochrome-conjugated monoclonal antibodies specific for cell surface antigens.
Staining cells with a Lyse/No-Wash protocol
Find procedures for whole blood specimen collection and preparation, including step-by-step instructions for lysing and staining. Use this method to detect cells bearing specific membrane antigens.
Explore four protocols using different methods (detergent method, mild or harsh alcohol method or harsh detergent method) for the activation of cell signaling molecules in mouse splenocytes or thymocytes.
Indirect Staining Mononuclear Cells
The indirect method is used to enhance the fluorescence signal and also to facilitate multicolor staining of human cells when direct conjugated reagents are not available. Find the procedure for specimen collection, preparation and staining of mononuclear cells.
Explore four protocols using different methods (detergent method, mild or harsh alcohol method or harsh detergent method) for the activation of cell signaling molecules in human peripheral blood mononuclear cells.
Explore four protocols using different methods (detergent method, mild or harsh alcohol method or harsh detergent method) for the activation of cell signaling molecules in human whole blood samples.
Reducing non-specific staining with Fc Block
Learn how antibody binding to Fc receptor can cause background staining and how to use the BD Fc Block™ Reagents in immunophenotyping of mouse or rat leucocytes..
Find the procedure for specimen collection, preparation and direct immunofluorescence staining of mononuclear cells..
Cell Surface Staining of Human PBMCs and Suspension Cell LinesD™ Phosflow Protocols for Human Whole Blood Samples
Find a step-by-step procedure for cell surface staining of human peripheral blood mononuclear cells and cell lines.
Find the optimal protocol for detecting intracellular signaling molecules from adherent cells using BD Phosflow™ Reagents.
Find answers to most commonly asked questions on intracellular flow cytometry and use of BD Phosflow™ Reagents..
Find the complete procedure for TCR stimulation of peripheral whole blood cells using BD Phosflow buffers.
Choosing the Right Fix Perm Kits
Choosing the Right Fix Perm Kits Cell Fixation/Permeabilization Kits for Intracellular Cytokine Analysis
Explore three cell fixation/permeabilization kits to simplify the preparation of cells for intracellular staining of cytokines.
The selection of an appropriate isotype control is essential for every flow cytometry experiment. Find procedures for determining the isotype of your primary antibody, for searching for your isotype control and for using your isotype control.
Find detailed information on electronic compensation for fluorochrome spectral overlap during flow cytometric analysis of multi-color immunofluorescence staining. We also offer a detailed procedure for setting compensation for multi-color flow cytometric analysis.
Escapees are aggregates of cells (including positively labeled cells) that are excluded from the flow cytometric analysis gate, thus altering the results of the analysis. See recommendations to decrease the number of escapees.
Intracellular Cytokine Staining Protocol
Get information on stimulation of cells, appropriate cultures for generating human, mouse and rat cytokine producing cells and describes a protocol for multicolor staining of intracellular cytokines and cell surface antigens. It also offers an alternative protocol for the activation and intracellular staining of whole blood.
Explore the complete procedure for cytokine stimulation of peripheral whole blood cells using BD Phosflow buffers.
Immune Cell Activation
Explore ways to activate human and mouse T cells. You can also find information on cultures for generation human, mouse or rat cytokine-producing cells.
Find protocols for fixing and permeabilizing cells using various BD Biosciences products, such as BD Cytofix™ Kits, BD Cytoperm™ Kits, BD GolgiStop™ and BD GolgiPlug™ Kits.
Downloadable BD Biosciences Protocols
Generation of Human IL-17–Producing Cells (Th17-like Cells) In Vitro Live Cell Discrimination and Measurement of Phosphorylation and Cell Surface Markers in Thawed and Activated Human PBMCs