The Protein Man's Blog | A Discussion of Protein Research

Lytic enzymes have a fairly prominent role in protein and DNA extraction. There are multiple enzymes that can be potentially used for lysis, depending on the applications and your lab's unique needs. Understanding lytic enzymes is essential to the process of successful protein extraction. Though physical lysis has been used in the past, it's no longer feasible -- and it's always important that lysis be completed with the proper lytic enzymes, to yield the most valuable results. 

Proteins are highly heterogeneous, complex bio macromolecules consisting of one or more long chains of amino acids. Proteins or peptides fold up to form secondary and tertiary structures, and associate with other protein subunits to form quaternary structures. Proteins are structurally and functionally different from each other and require distinct surrounding environment for their stability and activity. Proteins are susceptible to degradation, denaturation and precipitation when taken out of their native environment.

Carbohydrate affinity chromatography is a method of choice for purification of glycoproteins, lectins and other carbohydrate metabolite proteins. This affinity chromatography uses carbohydrate ligands such as carbohydrates, glycoproteins and carbohydrate matrices for purification. The matrix needs to be activated for covalent immobilization of the ligand. Various methods used for the coupling of ligands depending upon the activating reagents are as follows:

Gel filtration is a method of separating molecules by size. Among similar techniques, gel filtration is known as being a very simple and gentle procedure. When it comes to desalting and buffer exchange, gel filtration is often the preferred method as it can achieve better separation without any complex processes. Gel filtration is commonly used for chromatography, with the only negative being that it is very difficult to get high resolution results.