Molecular Probes™ Fluorescent Protein Labeling Kits

Description

• Ready to use in just 90 minutes, our protein labeling kits include an easy-to-use, pre-packed spin column for rapid dye removal and typical recovery greater than 85%
• Each kit contains enough reagent for 3–5 protein conjugation reactions
• Our kits provide better results due to lower background fluorescence, less nonspecific binding, and easier workflows for protein conjugation with 16 different fluorophores
• Fluorescently label up to 1 mg of protein per reaction (three reactions per kit) Label 0.5–3 mg per reaction with DSB-X Biotin Protein Labeling Kit (five reactions per kit) Labeled proteins ready to use in 90 min
• (∼15 min
• hands-on time) Rapidly purify proteins by quickly removing unbound dye using pre-packed Zeba Dye and Biotin Removal Spin Columns (7K MWCO, Cat
• No
• A44299) for >85% recovery Includes detailed instructions for determining degree of labeling (DOL) Each protein labeling kit contains everything you need to perform 3-5 separate labeling reactions and purify the resulting conjugates
• The reactive dye has either a succinimidyl ester (SE) or a tetrafluorophenyl (TFP) ester moiety that reacts efficiently with primary amines of proteins to form stable dye–protein conjugates
• Each of the vials of reactive dye provided in the kit is sufficient for labeling 1 mg of a variety of purified proteins, including growth factors, cytokines, nanobodies, enzymes, cell-adhesion molecules, and antibodies
• Direct labeling with fluorophores allows multiple primary antibodies of the same isotype (derived from the same species) to be used in the same experiment
• Stabilizing proteins such as BSA should be removed from the sample before labeling
• The different protein labeling kits Blue-fluorescent Alexa Fluor 350 —excitation and emission maxima of 346/442 nm Green-fluorescent Alexa Fluor 488 – excitation and emission maxima of 494/519 nm; excited using a 488 nm argon laser line and detected under standard FITC/Cy2 filters Yellow-fluorescent Alexa Fluor 532 – excitation and emission maxima of 530/554 nm; excited using a 532 nm Nd:YAG laser line and detected under standard Rhodamine 6G filters Orange-fluorescent Alexa Fluor 546 – excitation and emission maxima of 554/570 nm; excited using a 543 nm He-Ne laser line and detected under standard TRITC/Cy3 filters Orange-fluorescent Alexa Fluor 555 – excitation and emission maxima of 555/565 nm; excited using a 543 nm He-Ne laser line and detected under standard TRITC/Cy3 filters Orange-red-fluorescent Alexa Fluor 568 – excitation and emission maxima of 577/603 nm; excited using a 568 nm Kr laser line and detected under standard Rhodamine Red/Cy3.5 filters Red-fluorescent Alexa Fluor 594 – excitation and emission maxima of 590/617 nm; excited using a 594 nm Kr or He-Ne laser line and detected under standard Texas Red filters Far-red-fluorescent Alexa Fluor 633 – excitation and emission maxima of 632/647 nm Far-red-fluorescent Alexa Fluor 647 – excitation and emission maxima of 650/668 nm; excited using a 633 or 635 nm Kr or He-Ne laser line and detected under standard APC/Cy5 filters
• Far-red-fluorescent Alexa Fluor 660 – excitation and emission maxima of 663/690 nm Near-IR-fluorescent Alexa Fluor 680 – excitation and emission maxima of 680/700 nm Green-fluorescent Fluorescein-EX – excitation and emission maxima of 494/518 nm Oregon Green 488 – excitation and emission maxima of 496/524 nm Pacific Blue – a violet light excitable dye with an excitation and emission maxima of 410/455 nm Texas Red-X – excitation and emission maxima of 595/615 nm DSB-X Biotin – Conjugates can be reversibly bound to biotin-binding proteins such as streptavidin or avidin
• The concentration (mg/mL) of the DSB-X Biotin-labeled antibody preparation can be determined by measuring the absorbance of the dialyzed sample at 280 nm and dividing this value by 1.3 or 1.4 when measured in solution in a cuvette with a 1-cm pathlength
• DSB-X Biotin does not absorb significantly at 280 nm
• For labeling smaller amounts of antibodies (∼100 μg), we recommend our antibody labeling kits
• Please review the protein labeling kit user manual for more in depth information, protocols, molecular weights, and degree of labeling for each dye.