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Laboratory Protocols for Desalting Samples
Biological samples are commonly prepared in saline or other high salt buffers. High salt concentrations interfere with isoelectric focusing (IEF) of proteins during the first dimension separation. Desalting and/or buffer exchange columns are a common preparatory step in sample clean-up before proceeding to the first dimension of a 2-D gel. These columns contain a resin that removes salts, dye terminators, unincorporated nucleotides and other extremely low molecular weight molecules; also allowing for the reconstitution of sample protein into an IEF compatible matrix. Our facility primarily uses Pierce Zeba Desalt Spin Columns or Bio-Rad Micro-Spin Chromatography Columns for desalting and buffer exchanging of samples. Check the manufacturer's protocols for detailed instructions and ordering information to find the column size appropriate for your needs.
Pierce_Zeba_desalting_columns.pdf
BioRad_microspin_desalting_columns.pdf
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