The CF 597R Dye Antibody Labeling Kit is a high-performance fluorescent labeling system designed for conjugating antibodies with CF 597R, a far-red dye optimized for microscopy, flow cytometry, and other fluorescence-based applications. This labeling system provides high photostability, minimal photobleaching, and superior brightness, making it an ideal choice for researchers working in cell imaging and protein analysis.

Key Features and Applications

• Fluorescence Microscopy: The CF 597R dye is particularly well-suited for fluorescence microscopy due to its long-wavelength emission, reducing background noise and improving signal clarity. Learn more about fluorescence techniques from NIH and Harvard University.
• Flow Cytometry: The dye’s spectral properties align with many standard cytometers, making it a reliable choice for multiparametric analysis (FDA, Stanford).
• Western Blotting and ELISA: Antibody conjugates labeled with CF 597R dye exhibit excellent stability and sensitivity, crucial for protein quantification assays (NIH, CDC).
• Live and Fixed Cell Imaging: With a low degree of photobleaching, CF 597R enables long-term imaging in both live and fixed cell samples (University of California, National Institute of Standards and Technology).

Technical Specifications

• Excitation/Emission: 597 nm / 665 nm
• Quantum Yield: High quantum efficiency for improved signal-to-noise ratio (NIST).
• Hydrophilicity: Designed to minimize non-specific binding, ensuring cleaner results in biological assays (Johns Hopkins).
• Compatible Buffers: Works with common buffers like PBS, HEPES, and Tris, ensuring adaptability across protocols (CDC).

Advantages of CF 597R Over Other Dyes

Compared to conventional dyes like Alexa Fluor 594 and Cy5, CF 597R offers:

• Greater photostability for prolonged imaging sessions (NIH).
• Higher brightness, leading to enhanced resolution in microscopy (MIT).
• Minimal crosstalk in multicolor experiments, ensuring clearer fluorescence signals (FDA).

How the Labeling Kit Works

1. Activation of Antibody: The antibody’s amine groups react with CF 597R dye’s reactive moiety to form a stable covalent bond (NIST).
2. Purification: Free dye is removed via column filtration or dialysis to ensure only labeled antibodies remain (Harvard).
3. Validation: The conjugate is tested via spectrophotometry to confirm labeling efficiency and fluorescence intensity (Johns Hopkins).

Best Practices for Using CF 597R Dye

• Use freshly prepared buffers to maintain optimal reactivity (NIH).
• Avoid prolonged exposure to light to preserve dye stability (Stanford).
• Optimize dye-to-protein ratio to balance fluorescence intensity and antibody functionality (CDC).

Conclusion

The CF 597R Dye Antibody Labeling Kit is an advanced tool for biological imaging and flow cytometry, providing exceptional brightness, photostability, and compatibility across various applications. By following best practices and leveraging the latest protocols from leading research institutions, scientists can achieve highly reproducible and precise fluorescence-based analyses.

For further reading, visit resources from NIH, Harvard, and CDC.