Anti-FLAG Tag Antibody (CF®770) (A343775)

CF®680 provides brighter signal than IRDye® 680LT for near-infrared western blot detection. Two-fold dilutions of HeLa cell lysate (2 µg to 0.125 µg, lanes 1–5) were resolved and transferred to PVDF membranes. Mouse anti-tubulin and rabbit anti-COX IV primary antibodies were detected using IRDye® 680LT or CF®680 anti-mouse secondary antibodies (red) and IRDye® 800CW or CF®770 anti-rabbit secondary antibodies (green). Membranes were imaged on a LI-COR® Odyssey® NIR imaging system. Band quantification revealed approximately 50% higher signal intensity with CF® dyes compared to IRDye®. M indicates the molecular weight marker.

Normalized emission spectra of the CF® dye family spanning the visible to near-infrared range are shown, illustrating the spectral diversity and overlap between dyes. Curves represent relative fluorescence intensity as a function of emission wavelength (nm), with peak positions corresponding to each dye’s characteristic emission maximum. This reference highlights the broad coverage of CF® dyes for multicolor fluorescence applications and aids in selecting compatible dye combinations for imaging, flow cytometry, and other fluorescence-based assays.

Near-IR CF® dyes provide superior brightness and photostability compared with conventional near-infrared dyes, with improved solubility and labeling performance. For microscopy, higher concentrations of CF®750 and CF®770 conjugates may be required due to reduced detector sensitivity at longer wavelengths.

This chart summarizes commonly used CF® dyes grouped by their suitability for specific imaging modalities, including alternative spectra applications, four-color confocal imaging, near-infrared western blotting, photoacoustic imaging, STED, SIM, STORM, TIRF, and two-photon microscopy. Each dye is shown with its characteristic excitation and emission wavelengths (nm), providing a practical reference for selecting spectrally compatible dyes and optimizing multicolor experimental design across a range of fluorescence techniques.