With the use of flow cytometry in the clinical laboratory growing substantially in the past decade, facs-analysis.com is emerging as the top resource for all information related to flow cytometry.

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FACS Staining Protocol includes Antibody Staining, Direct staining, indirect staining and intracellular staining. In direct immunofluorescence staining, cells are incubated with an antibody directly conjugated to a fluorophore such as PerCP. This requires only one antibody incubation step and therefore eliminates the possibility of non-specific binding from a secondary antibody. Direct staining is advantageous during intracellular staining because large antibody-fluorophore complexes including secondary antibodies can become trapped and result in non-specific binding, or they may fail to enter the cell which results in no detection.

In indirect staining, the fluorophore conjugated secondary antibody detects the primary antibody which is unconjugated. Another available method is the avidin-biotin system, whereby a biotin-conjugated antibody is detected with fluorophore-labeled avidin.

The Intracellular Staining Protocol allows direct measurement of antigens (cytokines or transcription factors) present inside the cell cytoplasm or nucleus. In this procedure, the fixation and permeabilization of cells are required. Fixing cells will retain the target protein in its original cellular location, and will usually ensure better stability of soluble antigens and antigens with a short half-life. Detecting intracellular antigens requires cell permeabilization before staining, and antibodies should be prepared in permeabilization buffer to ensure the cells remain permeable.

Flow Cytometry Technical Resource Centre – FACS Analysis provides general experimental procedure optimized guide. The resource states that the primary requirement for all types of flow cytometry analysis is that the cells under analysis must be in a single-cell suspension. It is very important to obtain a single cell suspension to avoid clogging up the system with clumps. Peripheral blood mononuclear cells (PBMCs) isolated from whole blood through Ficoll gradient centrifugation, or RBC lysed whole blood, or non-adherent cultured cells are readily applicable for flow cytometry analysis.

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Founded in 1993 by famed histologist Steven Xia, Boster Bio is an antibody manufacturer specializing in high-sensitivity, high-specificity, ELISA kits and WB/IHC compatible antibodies. They have spent the last two and half decades perfecting the techniques and technology and their products have been well-cited in over 23,000 publications. Their antibodies are thoroughly validated for numerous applications including IHC, WB, ELISA and Flow Cytometry.

For more details & information about Flow Cytometry Technical Resource Centre, please click here: https://www.facs-analysis.com/.

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