I have to concentrate my enzyme for cellulase determination by ammonium acetate precipitation. the problem is due to unavailability of dialyzer. is there any alternate?
If you want to obtain the cellulase in an active form and determine its activity, start by ultra centrifugation of the precipitate, re-suspend the pellet in a buffer and perform gel filtration chromatography using the same buffer to elute the column. This will separate the protein from the residual salt and also fractionate the proteins in the extract by size.
Use ultrafiltration with a hollow fibre cartridge of 10 kDa cut off.
Alternatively, reconstitute protein precipitate in small volume of buffer and load on a Sephadex G-15 or G-25 column and collect early fractions containing cellulase, the salt ammonium acetate will be retained in the column matrix. This approach is referred as rapid dialysis.
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