AMCA-NHS | CAS 113721-87-2

AMCA-NHS is an N-hydroxysuccinimide ester of an AMCA (7-amino-4-methylcoumarin) fluorophore designed for covalent fluorescent labeling of primary amines. Its emission in the blue region makes it a useful choice for multicolor fluorescence workflows where spectral separation from common green and red dyes is required. The NHS ester format enables straightforward conjugation to proteins, peptides, and other amine-containing biomolecules used in microscopy, flow cytometry, and fluorescence-based assay development.

1. Protein And Antibody Labeling

AMCA-NHS is routinely used by protein chemistry and immunoassay teams to generate fluorescently labeled antibodies, antigens, and affinity reagents through covalent attachment to lysine residues and N-terminal amines. Researchers use the resulting AMCA conjugates to visualize binding interactions in fluorescence microscopy and to track reagent localization in cell-based binding studies. The AMCA fluorophore's blue emission supports multiplexing with many standard far-red or orange-red labels, helping reduce spectral overlap in imaging panels where multiple targets are assessed in parallel.

2. Flow Cytometry Fluorescent Label

AMCA-NHS is applied in flow cytometry reagent development to produce amine-reactive fluorescent conjugates for labeling cell-surface or intracellular targets after appropriate fixation/permeabilization workflows. Cytometry users value AMCA-NHS for constructing blue-channel-compatible dyes that can be combined with other fluorophores in multicolor panels, enabling simultaneous readout of marker expression and phenotypic gating strategies. In these workflows, the NHS ester chemistry is leveraged to prepare consistent fluorescent labeling reagents for antibody-based staining and fluorescence-based biomolecule tracking.

3. Fluorescent Bioassay Reagent Construction

AMCA-NHS is commonly incorporated into fluorescence assay development to prepare labeled detection reagents, such as fluorescently tagged proteins used as reporters in binding assays, wash-based immunoassays, and fluorescence readout formats. Assay developers use AMCA-NHS to introduce a stable fluorescent tag onto amine-containing components, supporting reagent tracking during protocol optimization and enabling quantitative fluorescence measurements in plate readers or imaging systems. The dye's spectral placement in the blue region helps fit into assay designs that require separation from assay autofluorescence and from longer-wavelength reporter channels.

4. Microscopy Multicolor Imaging

AMCA-NHS supports fluorescence microscopy workflows where blue-emitting labels are needed for cellular imaging, colocalization studies, or multicolor labeling schemes. Imaging labs use AMCA-NHS to prepare fluorescent conjugates of amine-containing biomolecules that can be introduced as staining reagents for target visualization, including labeled proteins used as probes in fixed-sample imaging and fluorescence localization experiments. The AMCA fluorophore's spectral characteristics help maintain channel separation when imaging multiple structures or biomolecules within the same specimen.