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.
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:
Tags: Protein Purification
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.
For the purposes of chromatography, fast flow agarose resin provides a stable, easy, and effective medium. Comprised of cross-linked agarose beads, fast flow agarose resin is able to improve upon both the speed and reproducibility of chromatography, offering good resolution and the ability to predictably scale up. Fast flow resins also have extremely good flow properties and superior loading capacity, making it ideal for faster separation cycles, better purification, and lessened dilution.
“Lectin” word originated from latin word “legree” which means “to select”. Lectins are proteins or glycoproteins that have at least one noncatalytic domain that binds specifically and reversibly to monosaccharides or oligosaccharides. Lectins whose sugar specificities are unknown are called hemagglutinins. Lectins are present in every organisms and are involved in biological functions such as adhesive, defence against pathogens, immunomodulatory and regulatory.
It's often necessary to determine the solubility of a chemical before you can begin working with it. Chemicals, especially those used in labs, are often concentrated and distilled, in order to be easier to use, transport, and store. But these highly concentrated chemicals will have to be properly reconstituted if they are to be used reliably. The solubility of a chemical is its ability to dissolve when presented with a certain solvent. When it comes to chemicals, it's often important to know how soluble the product is and which solvents are most appropriate for its applications.
There are a wide variety of ELISA substrates designed for different systems and detection methods. These ELISA substrates are designed to detect alkaline phosphatase (AP) and horseradish peroxidase (HRP). In addition to their sensitivity, other factors, such as cost and ease of use, may also factor in. Choosing the right ELISA substrates begins with determining the limits of detection required. From there, a general type of ELISA substrate can be selected and the specific substrate can be narrowed down. There are five major types of substrate available: PNPP, ABTS, OPD, TMB, and ONGP. Of these, TMB is one of the most versatile.
Cells are made up of recognizable structures that each serve their own purposes and have their own characteristics. Common subcellular markers are often used to distinguish cells from each other and to identify potential irregularities within the cellular structure. Though there are a multitude of subcellular markers available, a few of them are more common and more easily identified than others.