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

Different Types of Extraction Buffers and When to Use Them

Posted by The Protein Man on May 14, 2019 2:30:00 PM

Successful biochemical analysis heavily relies on the effective extraction of biologically active proteins from source materials (e.g., cell and tissue samples). Thus, you need to have an excellent working knowledge of your target protein(s) and use the appropriate extraction buffers for a given experimental design to ensure optimal protein recovery.

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Topics: Protein Extraction

Preventing Carbamylation When Using Urea as a Protein Solubilization Buffer

Posted by The Protein Man on May 7, 2019 2:30:00 PM

When it comes to solubilizing and denaturing proteins prior to isoelectric focusing and 2-D gel electrophoresis, most researchers choose urea. This is not surprising since this mild chaotropic agent can completely disrupt the interactions between protein molecules and increase the efficiency of protease activity in protein digestion while effectively preventing proteins from aggregating and/or precipitating. In addition, urea is also used for renaturing proteins from previously denatured samples.

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Topics: Protein Extraction

Tips for Preventing Protein Aggregation & Loss of Protein Solubility

Posted by The Protein Man on Jan 29, 2019 2:30:00 PM

Maintaining the stability and solubility of target proteins before, during and after purification is one of the biggest challenges researchers often run into. Proteins are extremely sensitive to solution conditions (i.e., pH and conductivity) and temperature. They also tend to aggregate at high concentrations required for structural studies. This can be a serious problem since protein aggregation can cause artifacts and hamper the biological activity of the target protein.

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Topics: Protein Extraction

How to Lyse Bacterial Cells

Posted by The Protein Man on Jan 15, 2019 2:30:00 PM

How do you break open bacterial cells in the lab? While the enzyme lysozyme is mainly responsible for lysing bacterial cells in nature, you can achieve the same effect by using particular enzymes, detergents, and chaotropic agents, and/or certain mechanical methods (e.g., sonication, repeated freezing and thawing, filtration, etc.).

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Topics: Protein Extraction

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