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

What is Protein Electrophoresis and When Should It Be Used?

Posted by Colin Heath on Nov 15, 2016 8:00:00 AM

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.

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A Derivatizing Agent for Glutathione Assays

Posted by The Protein Man on Nov 11, 2016 11:00:00 AM

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.  

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Topics: Apoptosis Assays

Glutathione Assays To Monitor Oxidative Stress

Posted by The Protein Man on Nov 10, 2016 9:53:28 AM

Monitor oxidative stress to determine the extent of cell damage using a colorimetric gluthathione assay.

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Topics: Cytotoxicity Assays

The Advantages of Loading Controls in Western Blotting

Posted by The Protein Man on Oct 18, 2016 8:00:00 AM

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. 

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