ATTO 465 NHS ester | CAS 1173020-88-6

ATTO 465 NHS ester is a reactive ATTO 465 fluorophore formulated as an N-hydroxysuccinimide (NHS) ester, enabling efficient covalent labeling of primary amines on proteins, peptides, and other amine-containing biomolecules. Its blue-emitting fluorescence makes it well suited for fluorescence microscopy, multicolor labeling workflows, and fluorescence-based assay development where robust dye conjugates are required.

1. Protein Conjugation Labeling

ATTO 465 NHS ester is widely used in protein labeling workflows to generate fluorescent protein conjugates for imaging and quantitative fluorescence readouts. Researchers typically apply it to antibodies, enzymes, and protein scaffolds where lysine residues and N-termini provide reactive primary amines for stable covalent attachment. The resulting ATTO 465 conjugates are commonly incorporated into binding studies, reagent development for fluorescence assays, and microscopy experiments that require a bright, blue spectral channel.

2. Fluorescence Microscopy Staining

ATTO 465 NHS ester supports fluorescence microscopy staining of fixed or prepared biological samples by covalently tagging amine-rich targets such as extracellular proteins, cell-surface proteins, or labeled biomolecular probes used in cellular imaging. In practice, it is used to build fluorescent labeling reagents for confocal and epifluorescence imaging, including multicolor experiments where ATTO 465 provides a distinct blue emission band. Conjugate-based staining workflows benefit from the NHS ester's chemistry-driven specificity for primary amines, helping reduce variability associated with noncovalent dye adsorption.

3. Flow Cytometry Labeling Reagents

ATTO 465 NHS ester is used to prepare fluorescent labeling reagents for flow cytometry, especially when amine-containing biomolecules such as antibodies, affinity reagents, or protein tags need to be converted into stable fluorescent conjugates. By targeting primary amines, the dye can be incorporated into antibody conjugates used for cell-surface or intracellular labeling strategies in assay development and phenotyping studies. The ATTO 465 emission is typically leveraged as a blue-channel fluorophore to support multiplex panels and consistent signal generation across labeling batches.

4. Fluorescent Bioassay Development

ATTO 465 NHS ester is commonly employed in fluorescence assay development to create labeled reagents used as detection components, tracers, or standards in plate-based formats. Researchers use ATTO 465 conjugates to monitor binding events, biomolecular interactions, or reagent performance in fluorescence readouts where covalent dye attachment to amine-containing partners improves workflow reproducibility. The NHS-ester labeling step is particularly useful when downstream assay components must remain fluorescent under repeated washing or buffer exchanges.