Proteinase K ( Lyophilized powder)

Proteinase K

⇓ Protocol

Proteinase K is a powerful and versatile enzyme widely used in molecular biology for its ability to digest a variety of proteins. Here’s a detailed overview:

What is Proteinase K?

this product is a serine protease that cleaves peptide bonds adjacent to the carboxyl group of aliphatic, aromatic, or hydrophobic amino acids. It is derived from the fungus Tritirachium album and is known for its robustness and broad substrate specificity.

Key Features :

• High Activity:  remains highly active over a wide range of temperatures (20-65°C) and pH levels (pH 4-12).

• Stabilit: It is stable in the presence of detergents (e.g., SDS) and denaturants (e.g., urea).

• Versatility: Effective in a variety of buffer systems and is resistant to autolysis.

Applications:

• DNA and RNA Extraction: Proteinase K is used to digest proteins during nucleic acid extraction, ensuring high purity and yield of DNA or RNA by removing contaminating proteins.

• Protein Digestion: Employed to digest proteins in cell lysates, tissues, and other biological samples for subsequent analysis.

• In Situ Hybridization: Used to permeabilize cells and tissues to allow probe access during hybridization procedures.

• Enzymatic Modifications: Inactivates nucleases (RNases and DNases) during sample preparation to protect nucleic acids from degradation.

Storage and Handling:

• Storage:Its typically stored as a lyophilized powder at -20°C. Once reconstituted, aliquots can be stored at -20°C for long-term use.

• Handling: Avoid repeated freeze-thaw cycles to maintain enzyme activity.

Proteinase K is a crucial enzyme in many molecular biology workflows due to its ability to effectively degrade a wide range of proteins under various conditions, facilitating the preparation of high-quality nucleic acid samples.

Applications

1. DNA and RNA Extraction:

   • Purpose: Ensures the removal of proteins and nucleases that can degrade nucleic acids.
   • Process: During extraction, Proteinase K is added to the lysis buffer to digest proteins, allowing for the purification of high-quality DNA or RNA.

2. Tissue Preparation:

   • Purpose: Prepares tissue samples for subsequent analysis or hybridization procedures.
   • Process: Proteinase K permeabilizes tissues, making them more accessible to probes or other reagents.

3. Histological Studies:

   • Purpose: Facilitates the study of tissues by breaking down extracellular matrix proteins.
   • Process: Proteinase K is used to digest protein components, making it easier to study tissue architecture and cellular components.

4. Proteomic Research:

   • Purpose: Investigates protein composition and interactions.
   • Process:  digests proteins in samples, allowing for analysis of peptides and protein fragments.