General Experimental Procedure

1. ========== Team Boster strives to provide best service and has earned the trust of researchers worldwide. All products are covered by the Boster Quality Guarantee, so each will work as advertised or your money back. Boster product lines are actively expanding with new antibodies and ELISA kits every month. Collaborations are always welcome! 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.

2. The Intracellular Staining Protocolallows 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 providesgeneral experimental procedureoptimized 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. About Facs About Facs- -Analysis Analysis 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/. ### Contact Details  Flow Cytometry Technical Resource Center 362 52nd Street, NYC, New York 10259 +357 720 1443 orders@bosterbio.com