The product named "Factory Supply Proteinase K CAS 39450 - 01 - 6 With Best Price" is Proteinase K. Proteinase K is a serine protease, and its CAS number is 39450 - 01 - 6. It is widely used in molecular biology and biochemistry research. This enzyme has high activity and stability under a wide range of conditions, including high - temperature and high - salt environments. The protease is produced by the factory with a guarantee of high quality and a competitive price. It is usually provided in a powder form, which can be easily dissolved in appropriate buffers for use. The specific activity of the proteinase K may vary according to different production batches, but it generally has a relatively high proteolytic ability.
1. **DNA and RNA Isolation**: Proteinase K is commonly used in the isolation of genomic DNA and RNA from biological samples. It can degrade proteins that are bound to nucleic acids, such as histones in chromatin, which helps to release pure DNA or RNA. For example, when extracting DNA from tissue samples, proteinase K is used to break down the proteins in the cell membrane and nuclear membrane to expose the DNA.
2. **In Situ Hybridization**: In in - situ hybridization experiments, proteinase K is used to permeabilize cells and tissues. This treatment allows probes to enter cells more easily and hybridize with their target nucleic acids, improving the efficiency and accuracy of the experiment.
3. **Protein Digestion**: In proteomics research, proteinase K can be used to digest proteins into peptides for further analysis, such as mass spectrometry analysis. It can specifically cleave peptide bonds at the carboxyl side of aliphatic, aromatic, or hydrophobic amino acids.
1. **Reconstitution**: First, reconstitute the powder - form Proteinase K in a suitable buffer. A common buffer is Tris - HCl buffer with a pH of around 7.5 - 8.0. The concentration of the reconstituted solution can be adjusted according to the specific experimental requirements, usually in the range of 1 - 20 mg/ml.
2. **Addition to the Reaction System**: Add the appropriate amount of proteinase K solution to the sample or reaction mixture. The amount depends on the nature and quantity of the sample. For example, when extracting DNA from a small amount of cells, 10 - 50 μl of a 20 mg/ml proteinase K solution may be sufficient for a 500 - μl reaction volume.
3. **Incubation**: Incubate the sample with proteinase K at an appropriate temperature. The optimal temperature for the activity of proteinase K is around 50 - 60°C, and the incubation time can range from 30 minutes to several hours, depending on the complexity of the sample and the digestion requirements.
1. **Case 1: DNA Extraction from Blood Samples**
A research team was extracting genomic DNA from blood samples for genetic analysis. They first collected 500 μl of whole blood. Then, they added 100 μl of proteinase K solution (20 mg/ml) to the blood sample, along with 500 μl of lysis buffer. The mixture was incubated at 55°C for 2 hours. After incubation, the proteins were effectively degraded, and the DNA was released. By subsequent purification steps, high - quality genomic DNA was obtained, which was suitable for downstream PCR and sequencing experiments.
2. **Case 2: Protein Digestion in Proteomics**
In a proteomics project, a scientist wanted to analyze the protein composition of a cell lysate. The scientist added 50 μl of proteinase K solution (10 mg/ml) to 500 μl of cell lysate. The mixture was incubated at 50°C for 3 hours. After digestion, the resulting peptides were analyzed by liquid chromatography - mass spectrometry (LC - MS). The results showed that the proteinase K effectively digested the proteins in the cell lysate, providing accurate peptide information for protein identification.
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