![]() Blue fluorescent dyes in particular such as CF®350, CF405S® and CF®405M have relatively lower fluorescence and photostability. Some fluorescent dyes photobleach more rapidly than others.Using mounting medium with antifade, such as EverBrite™ Mounting Medium.Use IgG-free BSA or fish gelatin for blocking to avoid cross-reactivity of secondary antibodies with immunoglobulins in the blocking agentįluorescence is photobleaching during microscopy.If using anti-goat or anti-bovine secondary antibodies, avoid blocking buffers with milk, goat serum, or BSA.Secondary antibody is not compatible with serum proteins used for blocking If the binding site is not known, perform intracellular staining to determine if target is localized inside the cell.Check whether the antibody immunogen or epitope is in an extracellular domain of the target protein, or is validated for cell surface staining. Even if the protein of interest is localized to the cell surface, your antibody may bind a cytoplasmic domain inside the cell. ![]()
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