Goat F(ab')2 Anti-Alpaca IgG, VHH domain + Llama IgG, VHH domain Antibody (CF®680) - Highly Cross-Adsorbed (A344044)

Comparison of microtubule imaging using conventional wide-field microscopy and STORM using a CF®680 dye conjugate. Images courtesy of Sam Kenny and Professor Ke Xu, College of Chemistry, University of California, Berkeley.

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.

CF®680R is highly photostable. Jurkat cells were stained with mouse anti-CD3 antibody and goat anti-mouse conjugates of the dyes shown. Cells were exposed to continuous mercury arc lamp excitation with a Cy®5 filter set. Images were captured every 10 seconds for 5 minutes; mean fluorescence was normalized to time 0.