The bicinchroninic acid (BCA) assay, also known as the Smith assay, is a biochemical assay used to determine the total concentration of protein in a solution. Due to its ability to provide accurate determination of protein concentration and its compatibility with most protein sample types, protein laboratories around the world prefer the BCA assay over any other detergent-compatible assays.
The Protein Man's Blog | A Discussion of Protein Research
We recently reviewed the advantages of total protein membrane stains as a loading control for Western blotting and demonstrated how they were an improvement on routinely used housekeeping genes.
Tags: Western Blotting
Western blotting is a very popular and sensitive protein detection system that is routinely used for multiple areas of protein research. The sensitivity of this detection system continues to improve with increasingly sensitive enhanced chemiluminescence systems and the increasing popularity of IR (infra-red) fluorophores and IR imaging systems:
Due to its high sensitivity, wide dynamic range, and high signal-to-noise ratio, enhanced chemiluminescence or ECL is considered as one of the most popular detection methods for a variety of western blotting applications in most protein laboratories around the world. This method also proves to be very useful in the quantification of a wide variety of biological materials such as cells, proteins, RNA, DNA, and a host of other analytes.
Tags: Western Blotting
Protein quantitation or estimation assays are widely used for determining protein concentration and are considered to be one of the most widely used methods in life science research. Estimation of protein concentration is necessary in protein purification, electrophoresis, cell biology, molecular biology, and other research applications.
Among all the protein staining techniques available today, most researchers around the world prefer using Coomassie dyes (also known as Coomassie brilliant blue) in visualizing electrophoresed proteins. There are a number of good reasons why they do. Here are some of them:
Tags: Protein Detection
Proteins separated by gel electrophoresis can be visualized using different staining procedures, including Coomassie stains, silver stains and fluorescent stains. Among these staining techniques, silver staining undoubtedly offers the highest sensitivity. However, it also involves a complex and relatively time-consuming protocol, has a low linear dynamic range and reproducibility, and is not compatible with mass spectrometry.
There are certain things that you should not do when performing dialysis in the laboratory. To ensure that all unwanted small molecular weight substances (such as reducing agents, non-reacted crosslinking or labeling reagents, and preservatives) are removed and facilitate a more efficient buffer exchange for macromolecular samples such as proteins, here are some of the things you need to avoid at all cost.
Tags: Sample Clean Up
lSodium dodecyl sulphate-polyacrylamide gel electrophoresis or SDS-PAGE is commonly used in protein aboratories for separating proteins based on their molecular weights. This is accomplished by taking advantage of their differential rates of migration through a sieving matrix (the acrylamide or agarose gel) in response to an applied electrical field.
While protein assays have a variety of uses in life science research, there is no single assay method that is suitable for all applications. Despite all the advancements in modern science, a protein assay method that is not affected by any non-protein components or by the differences in protein composition does not exist. For this reason, protein laboratories find it necessary to have more than one type of protein assay for research applications.
Tags: Protein Estimation