Visible Fluorescent Protein Labeling Kits

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Visible fluorescence protein labeling kits can be used to label antibodies and other proteins for applications such as flow cytometry, microscopy, immunohistochemistry, and other applications where fluorophore-conjugated antibodies are required.

The visible dyes included in these kits are activated with N-hydroxysuccinimide (NHS) esters, which is the most commonly used reactive group for labeling proteins. NHS esters react with primary amines, forming stable, covalent bonds.

VRDye labeling kits are based on a simple conjugation protocol that uses fast, spin column cleanup with resulting purity equivalent to purification by dialysis. Raise the pH of the preservative-free protein solution with the provided buffer, combine it with the appropriate amount of water-soluble amine-reactive dye, and separate the labeled conjugate from the free dye using the included spin columns. Labeling and purification are complete in approximately 2 hours.

Properties of Visible Fluorescence NHS Ester Dyes

Dye Absmax Emmax ε (M-1cm-1) MW (g/mole) CF*
IRDye 650 648 655 230,000 1,097 0.03
VRDye 549 550 575 150,000 1,040 0.07
VRDye 490 491 515 73,000 1,011 0.11

*CF is a correction factor for the absorbance of the dye at Absmax to the absorbance of the protein at 280 nm.

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