Resolution of Protein Disulphide-Isomerase Activities by Covalent Chromatography (CAT#: STEM-ACM-0215-CJ)

Covalent chromatography is a method for separation of molecules, based on formation of reversible covalent bonds between functional groups in molecules and complementary structures on a stationary solid phase. Covalent chromatography thus involves a synthetic step by which a solute is covalently immobilized to a solid support—the chemisorbent— later followed by chemical cleavage and regeneration of the sorbent.

Biochemical

1. Preparation of Column: The column is loaded with solid support such as sepharose, agarose, cellulose etc.. Ligand is selected according to the desired isolate. Spacer arm is attached between the ligand and solid support.
2. Loading of Sample: Solution containing a mixture of substances is poured into the elution column and allowed to run at a controlled rate.
3. Elution of Ligand-Molecule Complex: Target substance is recovered by changing conditions to favor elution of the bound molecules.

• Sample: Plants; Natural Food; Protein; Drug; Pollutants; Blood; Saliva; Serum; Plasma; Antibodies; Viruses & More
• Equipment: Agarose; Silica gel; Aluminium oxide; Acrylate; Organic polymers; Wash Buffer
• (Optional): Ligand; Spacer arm; Column

1. This approach is particularly popular for purifying enzymes and proteins. Dye-ligand adsorbents are desirable because they are affordable, readily available, and simple to immobilise. These adsorbents can be utilised for analytical, preparative, and large-scale research.
2. The dye-ligand affinity approach for pharmaceuticals has been discontinued due to leakage and toxicity issues.