What is Sepharose chromatography?

Sepharose is a tradename for a crosslinked, beaded-form of agarose, a polysaccharide polymer material extracted from seaweed. Its brand name is a portmanteau derived from Separation-Pharmacia-Agarose. A common application for the material is in chromatographic separations of biomolecules.

How does Q Sepharose column work?

The charged group of Q-Sepharose is a quarternary amine which carries a non- titratable positive charge. This matrix can be used at alkaline pH values at which the positive charge of the DEAE group would have been titrated. The charged group of S-Sepharose is the sulphonyl group (-SO3¯).

Is ion exchange chromatography affinity chromatography?

In ion exchange chromatography, molecules are separated according to the strength of their overall ionic interaction with a solid phase material (i.e., nonspecific interactions). By contrast, affinity chromatography (also called affinity purification) makes use of specific binding interactions between molecules.

What is the composition of Sepharose?

Chemical stability For daily use, Sepharose High Performance media are stable in all common, aqueous buffers, 1 M NaOH, denaturing agents (8 M urea, 6 M guanidine hydrochloride), 70% ethanol, 1 M acetic acid, 30% acetonitrile and with additives such as non-ionic detergents.

What is Protein A Sepharose?

Protein A Sepharose® beads are prepared by covalently coupling recombinant Protein A to 6% cross-linked Sepharose® beads. Protein A is a genetically engineered protein containing five IgG-binding regions of native Protein A.

How does ion exchange chromatography work?

Ion-exchange chromatography retains analyte molecules on the column based on coulombic (ionic) interactions. The ion exchange chromatography matrix consists of positively and negatively charged ions. Essentially, molecules undergo electrostatic interactions with opposite charges on the stationary phase matrix.

What is DEAE Sepharose fast flow?

DEAE Sepharose Fast Flow is a weak anion exchange chromatography resin (medium) for fast protein purification. DEAE Sepharose Fast Flow is a weak anion exchange chromatography resin (medium) for fast protein purification.

What is DEAE-Sepharose?

Diethylaminoethyl-sepharose (DEAE-sepharose) is a strong anion exchange column, where DEAE covalently binds to sepharose.

How should I pack my Sepharose fast flow media?

Ideally, Sepharose Fast Flow media are packed in XK columns in a two-step procedure: Do not exceed 0.5 bar (0.05 MPa) in the first step and 1.5 bar (0.15 MPa) in the second step.

What is the size of the Blue Sepharose CL-6B column?

A 36-ml packed Blue Sepharose CL-6B (Amersham Pharmacia Biotech) column, 26 mm in diameter, is prepared according to the instructions of the manufacturer. The column is equilibrated with buffer D at a flow rate of 34 cm/hr.