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Specifications
| Appearance: | Yellow powder |
| Purity: | 99% min (HPLC) |
| Absorption spectrum: | Conform |
| Fluorescence spectrum: | Conform |
| Structural confirmation: | Absorption and fluorescence peaks at the correct wavelengths |
| Solubility: | Soluble in dimethyl sulfoxide or dimethyl formamide |
Applications
1. Bioconjugation and Fluorescent Labeling
The free carboxyl group allows efficient coupling to amines (e.g., in proteins, peptides, or nucleotides) using EDC/NHS chemistry to form stable amide bonds.
Used to prepare fluorescently labeled antibodies, enzymes, oligonucleotides, and other biomolecules for cell imaging, tracking, and assays.
2. Fluorescence Microscopy and Imaging
Emits strong green fluorescence (excitation ~494 nm, emission ~518 nm), making it ideal for live-cell and fixed-cell fluorescence microscopy.
Used in cellular localization studies, intracellular tracking, and co-localization experiments.
3. Flow Cytometry and FACS
Applied in fluorescent cell labeling for quantitative analysis of cell surface markers, receptor binding, and cell viability using flow cytometry.
4. Fluorescent Probes and Sensors
Incorporated into biosensors and molecular probes for detecting pH, ions, or enzymatic activity.
Often used in fluorescent indicator systems due to its sensitivity and photostability.
5. DNA and RNA Labeling
Covalently linked to oligonucleotides for use in qPCR, microarrays, and fluorescent in situ hybridization (FISH).
Ensures precise labeling for quantitative nucleic acid detection.
Benefits
1. Single Isomer Purity
Unlike 5(6)-carboxyfluorescein, 6-carboxyfluorescein provides single-isomer consistency, ensuring reproducibility in experimental results.
Reduces variability in labeling efficiency and downstream biological assays.
2. Strong and Stable Fluorescence
High quantum yield and brightness support sensitive detection in low-concentration systems.
Good photostability, suitable for long-term fluorescence imaging.
3. Biocompatibility and Solubility
Soluble in aqueous buffers and biologically compatible at low concentrations.
Minimal toxicity makes it ideal for live-cell applications.
4. Efficient Bioconjugation Chemistry
The carboxyl group reacts efficiently under mild conditions, enabling fast and stable attachment to a wide range of biomolecules.
5. Broad Utility Across Research and Diagnostics
Widely used in commercial diagnostics, clinical research, and academic science due to its performance and versatility.
Conclusion
6-Carboxyfluorescein (CAS 3301-79-9) is a highly effective, single-isomer fluorescent dye used in bioconjugation, microscopy, flow cytometry, and molecular diagnostics. Its defined structure, bright green fluorescence, and reactive carboxyl group make it a preferred tool for researchers requiring precise, reliable labeling of biomolecules. Compared to mixed-isomer versions, its purity enhances reproducibility and simplifies downstream analytical work.
