When proteins need to be segregated out from a sample, protein electrophoresis may be used. Protein electrophoresis utilizes a matrix and an electrical current in order to easily separate proteins without disruption. Protein electrophoresis is beneficial for a few reasons: it can be performed on a relatively small sample size, it is accurate, and it is simple. There are a few different methods of protein electrophoresis, which may or may not involve a denaturing gel.
What is Protein Electrophoresis and When Should It Be Used?
4-Vinylpyridine as a derivatizing agent for free thiols and its application in Glutathione assay
Glutathione assay involves quantification of reduced Glutathione (GSH) and Oxidized Glutathione (GSSG) in various organisms or in various tissues, blood samples, plasma, serum or cultured cells. Free thiols such as GSH can be detected by their property of relatively high reactivity compared to other biological molecules. On contrary disulfides such as GSSG does not have any unique property that could be exploited for its quantification. Hence Glutathione disulfide (GSSG) quantification can be done after reducing it to its corresponding thiol (GSH). Thus the most widely adopted methodology for quantification of GSH and GSSG is determination of total GSH concentration, followed by alkylation to remove GSH and then reduction of GSSG and its quantification. GSH concentration in sample can be determined by subtracting glutathione disulfide concentration from total glutathione concentration.
Topics: Apoptosis Assays
Monitor oxidative stress to determine the extent of cell damage using a colorimetric gluthathione assay.
Topics: Cytotoxicity Assays
The Advantages of Loading Controls in Western Blotting
Loading controls serve as a vital method of verifying the results acquired through western blotting. Loading controls are antibodies that are used to detect proteins within samples. When western blots are used to determine the levels of protein expression in a sample, loading controls ensure that the results aren't due to loading or protein transfer errors. When using a loading control, the right type of control must first be identified.