Preparation of an Ion-Exchange Column Protocol Protocol for the preparation of ion-exchange chromatography column. Ion-exchange chromatography (IEC) can be used as a crude step in a protein purification scheme, or, with proper preparation, as a high-resolution step. If high resolution is desired, considerable care should be taken during column preparation, choice of IEC media, and column packing.
Purification of DNA Recovered Anion-exchange Chromatography Protocol Protocol for the purification of DNA recovered from agarose and polyacrylamide gels by anion-exchange chromatography. Fragments of DNA recovered from agarose gels are sometimes poor templates or substrates in subsequent enzymatic reactions. This problem can be solved by binding the DNA to a positively charged matrix, such as DEAE-Sephadex or DEAE-Sephacel, in buffers of low ionic strength. After washing the matrix, the DNA is eluted by raising the strength of the buffer.
Selection of an Ion Exchanger Protocol Protocol for selection of an ion exchanger: Determining the pI of a protein using isoelectric focusing. The choice of whether to use an anion or a cation exchanger should be based on knowledge of the stability of the protein, and the binding properties of the target protein and other molecules present.
Selection of an Ion Exchanger Protocol II Protocol for selection of an ion exchanger: Determining the pI of a protein using the titration curve method. The choice of whether to use an anion or a cation exchanger should be based on knowledge of the stability of the protein, and the binding properties of the target protein and other molecules present in the sample.
Selection of an Ion Exchanger Protocol III Protocol for selection of an ion exchanger: Determining the pI of a protein using the test tube method. The choice of whether to use an anion or a cation exchanger should be based on knowledge of the stability of the protein, and the binding properties of the target protein and other molecules present in the sample.
Selection of an Ion Exchanger Protocol IV Protocol for selection of an ion exchanger: Determining the pI of a protein using the Trial-and-Error method. The choice of whether to use an anion or a cation exchanger should be based on knowledge of the stability of the protein, and the binding properties of the target protein and other molecules present in the sample.
Separating Proteins Using Ion-Exchange Chromatography Protocol Protocol for separating proteins using ion-exchange chromatography. Protocol details the practical considerations of an ion-exchange chromatography (IEC) experiment. The choice of what type of ion exchanger to use, as well as the composition of the buffers used in this experiment, should be determined prior to beginning this protocol.
