Per the product technical bulletin, the labeling reaction is sufficient for 1 mg of protein at a concentration of 2–10 mg/mL, and the two-hour incubation step is at room temperature.
76508
Atto 647N Protein Labeling Kit
BioReagent, suitable for fluorescence
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1 KIT
€857.00
About This Item
UNSPSC Code:
12352108
NACRES:
NA.32
Recommended Products
product line
BioReagent
Quality Level
manufacturer/tradename
ATTO-TEC GmbH
fluorescence
λex 647 nm; λem 661 nm in 0.1 M phosphate buffer, pH 7.0 (recommended)
suitability
suitable for fluorescence
storage temp.
2-8°C
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General description
This kit contains sufficient amounts of reactive dye, buffers and protein purification sets for performing five labeling reactions (1 mg protein each) and for the subsequent purification of the labeled protein.
Legal Information
This product is for Research use only. In case of intended commercialization, please contact the IP-holder (ATTO-TEC GmbH, Germany) for licensing.
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Storage Class Code
10 - Combustible liquids
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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Articles
Protein labeling kits with Atto and Tracy dyes provide easy fluorescent labeling of purified proteins, enzymes, and antibodies.
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Is there an optimal volume of protein at c=2mg/ml (e.g. is it better to have 1 mg in 0.5ml PBS or 2mg in 1ml PBS). Incubation of 2h is performed on RT or +4C?
1 answer-
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Will this kit efficiently label protein on a much smaller scale of < 20ug of protein or at concentration below 2 mg/ml?
1 answer-
The dye should be reconstituted in 20 ul of DMSO and the total volume added to 1 mg of the antibody (at 2-10 mg/ml). If the amount of antibody is less, it needs to be at 2 mg/ml, and the amount of dye added to the antibody solution should be proportionally reduced. For example, if there is 200 ug of antibody (at 2 mg/ml), 4 ul of the DMSO solution of the dye should be added to the antibody. It's important for the user to use very accurate pipettes to avoid adding too much dye to the antibody, which could result in the labeling of the antibody in the antigen-binding site. If less protein is used, the amount of dye can be reduced. There is always an excess of dye to ensure labeling efficiency, and a part of the dye will be separated (as unbound dye) by a purification step afterward. The proteins need to have a minimum concentration due to the strong hydrolysis tendency of the NHS-functionality. If the protein solution is too dilute, the dye will be hydrolyzed before it reaches an amine-group of a protein.
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