Poor protein transfer can lead to either a weak signal or a lack of signal being observed during the process of western blotting. Though poor protein transfer does occur fairly often, it's also commonly disregarded as a potential cause for a failure. Consequently, it's an issue that should be considered first when trying to determine the cause of a poor signal.
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.
Topics: Protein Purification, Protein Extraction
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:
Topics: Protein Purification
