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Concentration Protein after fluorescent labeling

Concentration Protein after fluorescent labeling - Protein Forum

Concentration Protein after fluorescent labeling - Protein Forum


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Old 06-18-2009, 11:30 AM
Peggy Willemarck
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Default Concentration Protein after fluorescent labeling



Does anyone knows how the concentration of labeled protein with Alexa 647 can be determined? Normal methods such as OD280, Bradford and coomassie stain have to much interference with the dye to give a good concentrations.

Peggy








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Old 06-19-2009, 12:11 AM
DK
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Default Concentration Protein after fluorescent labeling

In article <[Only registered users see links. ].net >, Peggy Willemarck <[Only registered users see links. ]> wrote:

Bradford *is* Coomassie-based!


1. BCA assay is expected to not be affected by Fluorescent labels.
2. UV is pretty trivial, too: assuming you have a protein labeled at
near 100%, all you do is to add protein's and dye's extinction coefficients
to give epsilon of the label protein. Alexa's absorption at 280 is either
published somwhere or you can get free dye and measure it yourself.

DK
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