ATTO465 NHS ester | CAS 1300667-93-9

ATTO465 NHS ester is a reactive fluorescent dye for covalent protein labeling via NHS-ester chemistry, enabling attachment to primary amines on antibodies, enzymes, peptides, and other bioconjugation targets. Its visible excitation/emission profile makes it a practical choice for standard fluorescence microscopy and fluorescence readouts, including conjugate-based imaging and assay development workflows where robust labeling is required.

1. Antibody And Protein Labeling

ATTO465 NHS ester is widely used by antibody and protein engineering groups to generate fluorescently labeled immunoreagents for binding studies, target localization experiments, and reagent benchmarking. The NHS-ester functionality reacts with lysine residues and N-terminal amines, allowing straightforward preparation of fluorescent antibody conjugates and enzyme conjugates for downstream assays. Researchers typically use these labeled proteins in fluorescence microscopy and plate-based fluorescence assays to track binding, distribution, or relative abundance of protein targets under their experimental conditions.

2. Fluorescence Microscopy Staining

ATTO465 NHS ester supports fluorescence microscopy workflows where covalent labeling of fixed or prepared proteinaceous samples is needed for cellular imaging and biomolecular localization. Common use cases include labeling extracellular matrix components, labeling protein markers in immunostaining workflows, and preparing fluorescent conjugates for imaging studies that require stable attachment to amine-containing biomolecules. In microscopy and imaging reagent development, the dye's visible spectral window helps integrate labeled conjugates into multi-color experiments using standard filter sets and fluorescence imaging platforms.

3. Flow Cytometry Conjugates

ATTO465 NHS ester is used in flow cytometry reagent development to prepare amine-reactive fluorescent conjugates for labeling cell-associated proteins and other biomolecules present on cell surfaces or in prepared samples. Typical workflows involve generating fluorescent protein conjugates that can be incorporated into staining panels for fluorescence readout on flow cytometers, enabling quantitative comparison of labeled populations based on fluorescence intensity. The covalent NHS-ester labeling approach is particularly useful when stable labeling of protein reagents is required for consistent staining across experimental batches.

4. Fluorescent Bioassay Reagents

ATTO465 NHS ester is applied in fluorescence-based bioassay development where fluorescent protein conjugates serve as detection reagents or assay components. Researchers use dye-labeled proteins to build fluorescence readouts for binding assays, reagent performance testing, and assay optimization studies that rely on covalent attachment of the fluorophore to amine-containing assay components. This labeling strategy is also commonly used to prepare fluorescent standards and tracer reagents for method development in fluorescence spectroscopy and plate reader formats.