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"I have used a wide variety of secondaries and Jackson ImmunoResearch has consistently been the best. The fluorophores are bright and stable and their selective (x reactivity removed) secondaries have always shown species specificity in multiple labeling."Janet Duerr, Ohio University
Aminomethylcoumarin Acetate (AMCA) conjugates absorb light maximally around 350 nm and fluoresce maximally around 450 nm. For fluorescence microscopy, AMCA can be excited with a mercury lamp and observed using a UV filter set. Since blue fluorescence is not well detected by the human eye, AMCA-conjugated secondary antibodies should be used with the most abundant antigens in multiple labeling experiments. Ways of improving the visibility of AMCA include dark adapting the eyes, using fluorite instead of glass objectives, avoiding mounting media that absorb UV light (such as plastic-based media), and capturing photographic images with blue-sensitive film or CCD cameras. AMCA fades rapidly in conventional epifluorescence and confocal microscopy, and therefore it should be used with mounting media containing an anti-fading agent such as n-propyl gallate.
For flow cytometry, AMCA can be excited with a mercury lamp or with a water-cooled argon ion laser which emits some lines in the UV. AMCA has been used mostly for multiple labeling since there is minimal fluorescence overlap with green-fluorescing dyes and little or no overlap with longer wavelength-emitting fluorophores.
AMCA is available conjugated to:
|Excitation Peak (nm)||Emission Peak (nm)|
|Aminomethylcoumarin Acetate, AMCA||350||450|
Use the spectra viewer below to compare AMCA with other fluorophores, along with instrument specifics (laser/filter) for suitability in your assay.