AFDye 488 Antibody Labeling Kit

The AFDye 488 Antibody Labeling Kit provides all of the necessary reagents to perform labeling of small amounts of antibodies or other proteins (except IgM antibodies) with green-fluorescent AFDye 488 (Alexa Fluor® 488 equivalent). Simply mix your antibody with the reaction additive and pre-measured dye provided, followed by a brief incubation, and the conjugate is ready for simple purification or staining without further purification. The antibody will be covalently labeled with an average of 4-6 dyes per label molecule per antibody molecule. This kit is optimized for the labeling of 100 µg of antibodies per reaction at 1 mg/mL.

This kit utilizes 2,3,5,6-tetrafluorophenyl (TFP) esters instead of succinimidyl ester (SE or NHS) often used in conventional labeling kits. TFP is another type of carboxylic acid derivative that reacts with primary amines to form covalent amide bonds. The amine linkage bond is identical to the one formed by the reaction between primary amines and NHS esters or sulfo-NHS esters. However, in most cases, TFP ester displays much better stability toward hydrolysis in aqueous media resulting in more efficiency and better reproducibility in labeling of biopolymers. As a result of improved efficiency, very little of the non-reactive (hydrolyzed) dye is left in the labeling mixture, which allows staining without further purification.

AFDye 488 produces pH insensitive (pH 3-10), more photostable, and brighter protein conjugates compared to the previous generation of dyes (fluorescein/FITC). AFDye 488 labeled antibodies can be used for different applications, such as flow cytometry, fluorescent microscopy, ELISA, and Western blotting.

The AFDye 488 Antibody Labeling Kit can be used as direct, less expensive replacement of Alexa Fluor® 488 Antibody Labeling Kit

Alexa Fluor is a registered trademarks of Thermo Fisher Scientific.

Labeling Method: Covalent labeling

Labeling Target: Lys (primary amine)

Number of Labeling Reactions: 1

Abs/Em Maxima: 494/517 nm

Spectrally Similar Dyes: FAM, Alexa Fluor® 488, Atto™ 488, CF® 488A Dye, DyLight® 488