Today it is an established fact that Flow Cytometry is a powerful analysis tool in modern biology and life sciences. The applications are numerous and have contributed into many fields including virology, immunology, and molecular biology. The major advantages include the analysis of high cell numbers per sample combined with an increasing number of parameters. Based on the expression pattern of those parameters, it is possible to isolate specific cell populations for further analysis.

FACS-Analysis.com is technical resource centre for flow cytometry. FACS Antibody with single cells stained with fluorescent dye coupled monoclonal antibodies (or other bio reactive molecules) emit characteristic light signals, while passing one or more focused laser beams. These events can then be collected and analysed.

The core facility flow cytometry is equipped with various instruments and software modules to analyse and sort your samples. Flow Cytometry Antibody Staining Procedure includes direct staining, indirect staining and intracellular staining.

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. If the experiment will be staining with unconjugated purified antibody, there needs to be an additional step of staining with a fluorescent conjugated secondary antibody (indirect staining).

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. There is a wide range of conjugated antibodies available today, and with indirect staining, the choices of target proteins further increases.

In addition to the cell surface antigens, the intracellular staining procedure 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. FACS-Antibody team plan, perform and interpret your FACS (fluorescence activated cell sorting) experiments and cell sorting.

FACS Protocols:

Direct immunostaining of surface antigens
Indirect immunostaining of surface antigens
General immuno-staining procedure for intracellular antigens
Intracellular cytokine/phospho-immunostaining
In Vitro Cell Stimulation Reference Table
Dye efflux staining
DNA content or Cell cycle analysis

About FACS-Analysis.com:

As the flow cytometry technical resource center – Boster offers information about FACS principle, FACS Sample Preparation, FACS Protocols, FACS Optimization and more.

Website: https://www.facs-analysis.com/.

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