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| Name | Proteinase K |
| Source | enzyme from a fungus |
| Applications | DNA and RNA isolation, protein purification, tissue digestion |
| Molecular Weight | 28,900 Da |
| Optimal Temperature | 37-65°C |
| Optimal PH | 8.0-12.0 |
| Inhibition | inhibited by PMSF and metal chelating agents |
| Storage | stable at -20°C for long term storage |
| Specificity | broad substrate specificity for cleaving peptide bonds |
| Activity | highly active in the presence of denaturing agents like SDS |
| Purification | purified by chromatographic techniques |
| Sensitivity | sensitive to protease inhibitors |
FAQ
What is Proteinase K and how is it used in research applications?
Proteinase K is a broad-spectrum serine protease that is commonly used in molecular biology and biochemistry research. It is able to degrade proteins by cleaving peptide bonds, making it a valuable tool for a wide range of applications.
Why is Proteinase K preferred over other proteases for certain experiments?
Proteinase K is particularly useful in research because of its ability to remain active in the presence of denaturing agents such as SDS and urea. This makes it well-suited for applications that involve disrupting protein structures or working with complex samples.
How is Proteinase K typically used in nucleic acid extraction protocols?
Proteinase K is often used in nucleic acid extraction protocols to degrade proteins that may interfere with downstream applications such as PCR or sequencing. By adding Proteinase K to a sample, researchers can effectively remove protein contaminants and isolate high-quality nucleic acids.
What are some common troubleshooting issues associated with Proteinase K usage?
One common issue that researchers may encounter when using Proteinase K is inactivation of the enzyme due to high temperatures or inappropriate buffer conditions. It is important to carefully follow the manufacturer's instructions and optimize conditions to ensure optimal enzyme activity.
What are the storage and stability considerations for Proteinase K?
Proteinase K should be stored at -20°C or colder to maintain its stability. It is also important to avoid repeated freeze-thaw cycles, as this can lead to a loss of enzymatic activity. By properly storing Proteinase K, researchers can ensure its effectiveness and reliability in their experiments.
Proteinase K is a broad-spectrum serine protease that is commonly used in molecular biology and biochemistry research. It is able to degrade proteins by cleaving peptide bonds, making it a valuable tool for a wide range of applications.
Why is Proteinase K preferred over other proteases for certain experiments?
Proteinase K is particularly useful in research because of its ability to remain active in the presence of denaturing agents such as SDS and urea. This makes it well-suited for applications that involve disrupting protein structures or working with complex samples.
How is Proteinase K typically used in nucleic acid extraction protocols?
Proteinase K is often used in nucleic acid extraction protocols to degrade proteins that may interfere with downstream applications such as PCR or sequencing. By adding Proteinase K to a sample, researchers can effectively remove protein contaminants and isolate high-quality nucleic acids.
What are some common troubleshooting issues associated with Proteinase K usage?
One common issue that researchers may encounter when using Proteinase K is inactivation of the enzyme due to high temperatures or inappropriate buffer conditions. It is important to carefully follow the manufacturer's instructions and optimize conditions to ensure optimal enzyme activity.
What are the storage and stability considerations for Proteinase K?
Proteinase K should be stored at -20°C or colder to maintain its stability. It is also important to avoid repeated freeze-thaw cycles, as this can lead to a loss of enzymatic activity. By properly storing Proteinase K, researchers can ensure its effectiveness and reliability in their experiments.
