Stain Buffer (FBS)

Stain Buffer (FBS) can be used for the immunofluorescent staining of single-cell suspensions prepared from either lymphoid tissues, bone marrow, peripheral blood, or cultured cells. Stain Buffer (FBS) is useful for the dilution and application of fluorescent reagents as well as for the suspension, washing, and storage of cells destined for flow cytometric analysis. Based on previous reports of staining media, Stain Buffer (FBS) was formulated as a neutral pH (pH 7.4)-buffered salt solution (i.e., DPBS) that is supplemented with heat-inactivated (56°C, 30 minutes) fetal bovine serum (FBS) proteins. As such, Stain Buffer (FBS) is designed to maintain cell viability and maximize fluorescence signal intensities generated by pH-sensitive fluorochromes, e.g., fluoresceine isothiocyanate (FITC). In addition, Stain Buffer (FBS) contains the metabolic inhibitor, sodium azide (NaN3). NaN3 inhibits the potential redistribution of cell surface antigens (e.g., due to shedding or internalization) caused by antibody crosslinking. NaN3 (in combination with maintenance of cold ambient temperatures) thereby prevents the potential loss of fluorescent signal intensities generated by immunofluorescently-stained cells during subsequent flow cytometric analysis.

Stain Buffer (FBS) can be used for the immunofluorescent staining of single-cell suspensions prepared from either lymphoid tissues, bone marrow, peripheral blood, or cultured cells. Stain Buffer (FBS) is useful for the dilution and application of fluorescent reagents as well as for the suspension, washing, and storage of cells destined for flow cytometric analysis. Based on previous reports of staining media, Stain Buffer (FBS) was formulated as a neutral pH (pH 7.4)-buffered salt solution (i.e., DPBS) that is supplemented with heat-inactivated (56°C, 30 minutes) fetal bovine serum (FBS) proteins. As such, Stain Buffer (FBS) is designed to maintain cell viability and maximize fluorescence signal intensities generated by pH-sensitive fluorochromes, e.g., fluoresceine isothiocyanate (FITC). In addition, Stain Buffer (FBS) contains the metabolic inhibitor, sodium azide (NaN3). NaN3 inhibits the potential redistribution of cell surface antigens (e.g., due to shedding or internalization) caused by antibody crosslinking. NaN3 (in combination with maintenance of cold ambient temperatures) thereby prevents the potential loss of fluorescent signal intensities generated by immunofluorescently-stained cells during subsequent flow cytometric analysis.

Flow cytometry (Routinely Tested)

Aqueous buffered solution containing fetal bovine serum and ≤0.09% sodium azide.

Development References(5) 1. Cytokine/Chemokine Manual. Genes to Proteins to Cells. Application Manual. 2nd Edition.. :29-47. 2. Holmes K, Lantz LM, Fowlkes BJ, Schmid I, Giorgi JV. Preparation of cells and reagents for flow cytometry.. Curr Protoc Immunol. 2001; Chapter 5:Unit 5.3. (Biology). 3. Jackson AL, Warner NL. Rose NR, Friedman H, Fahey JL, ed. Manual of Clincial Laboratory Immunology, Third Edition. Washington DC: American Society for Microbiology; 1986:226-235. 4. Mishell BB, Shiigi SM. Barbara B. Mishell and Stanley M. Shiigi., ed. Selected methods in cellular immunology. San Francisco: Freeman; 1980:3-27. 5. Ohkuma S, Poole B. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc Natl Acad Sci U S A. 1978; 75(7):3327-3331. (Biology).