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antibody concentration for desalting/conjugating to minileak

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#1 laur



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Posted 05 April 2014 - 10:18 PM

Hi all,


I am very confused as to how much protein to use. I would like to desalt an antibody via gel filtration just in 10 ml sephadex G-15 column to prepare it for conjugation to some minileak beads. The person who stored the antibody said to use 2mg/ml before he went away.


The information that I have while going through lab books, I think (his hand writing is worse than my skill at calculating these matters ) the A280 is roughly 49.9 mg/ml and the volume was concentrated to 257 ul. I am assuming the extinction co-efficient is 1.4 mg-1 as it was clear that his total amount of protein is 9 mg. Then on the tubes in the freezer has 36 mg/ml in what looks like 10 ml aliquots. I am confused as to how much to use concentration wise and volume wise for the task.


Any ideas about this or how to approach these kinds of calculations in future would be most welcome.

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