HiLyte™ Fluor 647 acid, SE (Ex/Em=647 nm/ 679 nm) is an excellent fluorescent labeling dye that generates the protein conjugates that are slightly red-shifted compared to those of Cy® 5 dyes, resulting in an optimal match to filters designed for Cy®5 dyes. However, total fluorescence of the secondary antibody conjugates of HiLyte™ Fluor 647 dyes is significantly higher than that of Cy®5 conjugates supplied by other companies. Also, unlike Cy®5 dyes, HiLyte™ Fluor 647 dyes have very little change in absorption or fluorescence spectra when conjugated to most proteins, oligonucleotides and nucleic acids, thus yielding greater total fluorescence at the same degree of substitution. The HiLyte™ Fluor 647-protein conjugates are suitable for immunofluorescent staining, in situ hybridization, flow cytometry and other biological applications. Additionally, our in-house data indicated that HiLyte™ Fluor 647 is superior to Cy®5 in fluorescence polarization-based assays. The kit has all essential components for performing the conjugation reaction and for purifying the HiLyte™ Fluor 647-protein conjugate. The entire process takes only half an hour. The kit is able to perform three conjugation reactions with each reaction labeling up to 5 mg IgG.
Component A: HiLyte™ Fluor 647 SE Amino-reactive dye: 3 vials
Component B: Reaction buffer for pH adjustment of the conjugation reaction: 0.5 mL
Component C: Spin column to purify dye-protein conjugate: 3 pre-packed columns
Component D: DMSO solvent for preparing dye stock solution: 1 mL
Component E: 10x Elution buffer for eluting dye-protein conjugate: 30 mL
Storage & stability
Store all kit components at 4°C; Keep Component A away from light and protected from moisture