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

Why do I need a protease inhibitor?

Posted by Protein Man on Aug 28, 2015 10:00:00 AM

While proteolytic enzymes such as proteases and phosphatases play an important role in living cells and help ensure the survival of the organism, the mechanisms that regulate the tightly controlled cellular environment is disrupted during cell lysis. When this happens, these enzymes may start cleaving a variety of proteins that they would otherwise not touch in intact cells. This situation often leads to reduced recovery of total protein and biologically meaningless representation of protein activities.

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Using Protease Assays for Accurate Protease Detection

Posted by Protein Man on Aug 20, 2015 10:00:00 AM

Proteases are enzymes that facilitate proteolysis, or the breakdown of protein and peptide molecules into smaller polypeptides and/or amino acids. These enzymes do their job by cleaving the peptide bonds linking the amino acids together in the polypeptide chains.

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Tags: Protein Estimation

The Common Failures That can Occur When Using Dialysis Bags

Posted by Protein Man on Aug 11, 2015 10:00:00 AM

Dialysis is a solution-based separation technique used to facilitate the removal of small, unwanted compounds from macromolecules such as proteins, DNA, or polysaccharides through a semi-permeable membrane using the principle of selective diffusion. Although there are a lot of factors that affect the dialysis rate, the semi-permeable membrane is the single most important factor that can determine the success of the process.

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3 Protein Gel Stains and When You Should Use Them

Posted by Protein Man on Aug 6, 2015 10:00:00 AM

After the protein bands have been separated by electrophoresis, researchers perform a dye-binding or color-producing chemical reaction to visualize the proteins embedded within the pores of the polyacrylamide gel matrix. Some of the most popular in-gel detection methods include staining with Coomassie dye, metallic silver, and zinc.

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Tags: Protein Estimation

Why BCA Protein Assay is Overhyped

Posted by Protein Man on Jul 3, 2015 10:00:00 AM

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.

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Are Total Protein Membrane Stains Compatible With IR Imaging Systems?

Posted by Protein Man on Jun 29, 2015 2:36:00 PM

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.

What's best? Housekeeping genes or total protein for Western blot loading controls

 

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Tags: Western Blotting

What's best? Housekeeping genes or total protein for Western blot loading controls

Posted by Protein Man on Jun 29, 2015 2:34:00 PM

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:

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Tags: Western Blotting, Protein Detection

Enhanced Chemiluminescence: What is it?

Posted by Protein Man on Jun 29, 2015 10:00:00 AM

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.

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Tags: Western Blotting

How to Deal with Interfering Agents in Protein Estimation Assays

Posted by Protein Man on Jun 10, 2015 10:00:00 AM

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.

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Coomassie, Enhanced Coomassie, and Colloidal Coomassie: What’s the Difference?

Posted by Protein Man on Jun 4, 2015 10:00:00 AM

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:

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Tags: Protein Electrophoresis, Protein Detection

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Technical Handbooks

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