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

Protein Quantitation in the Presence of Reducing Agents and Detergents with UPPA™

Posted by The Protein Man on Jul 14, 2021 1:00:00 PM

 

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Topics: Protein Purification

Lets Talk About Inclusion Bodies

Posted by The Protein Man on Mar 23, 2021 2:00:00 PM

 

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Topics: Protein Purification, Western Blotting, Protein Electrophoresis, Protein Estimation, Sample Clean Up, Protein Concentration, Protein Fractionation, Protein Extraction, Buffers & Chemicals, Protein Detection

All about Proteinase K

Posted by The Protein Man on Mar 16, 2021 1:00:00 PM

Proteinase K is a serine protease, the presence of a catalytic triad characterizes serine proteases, the catalytic triad is a cluster of three amino acids that make the catalytic center and consists of serine, aspartic acid, and histidine amino acids, which can often vary but all of these enzymes have a nucleophile serine and the same catalytic mechanism. Proteinase K has a catalytic triad consisting of Ser 224, His 69, and Asp 39. The substrate recognition sites are made up of peptide chains, 99-104 and 132-136.

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Topics: Protein Purification, Molecular Biology

Detergents and Proteins, a Refresher

Posted by The Protein Man on Feb 10, 2021 3:00:00 PM

Detergents are amphipathic compounds with a nonpolar, hydrophobic tail and a polar, hydrophilic head group. Due to these structural features detergents tend to aggregate into structures called micelles at high enough concentration; arranging themselves with their hydrophobic tails pointed inwards and their hydrophilic heads pointed outwards. Detergents come in three types: ionic (cationic and anionic) and non-ionic. Non-ionic detergents aren’t generally used for gel electrophoresis due to their limited ability to break non-covalent interactions between protein residues and inability to impart a uniform charge onto the protein. Ionic detergents (typically anionic SDS) are used for gel electrophoresis as they are highly useful for protein solubilization, linearization and for establishing a uniform charge in preparation for gel electrophoresis.

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Topics: Protein Purification, Western Blotting, Protein Estimation, Detergents, Sample Clean Up, Protein Concentration, Protein Fractionation, Protein Labeling, Protein Extraction, Protein Detection

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