Code | CSB-AP361972TIQ |
Size |
Purchase it in Cusabio online store (only available for customers from the US) |
Image |
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Description |
Proteinase K, originally isolated from the mold Tritirachium album, is a serine protease with broad substrate specificity and relatively high proteolytic activity. It preferentially cleaves ester and peptide bonds adjacent to the C-termini of hydrophobic, aliphatic, or aromatic amino acids. CUSABIO's recombinant proteinase K is highly pure determined by SDS-PAGE and reached up to 95%. It is characterized by high purity, sterility, no bio-burden, and no presence of DNAse, RNAse, DNA, and RNA contaminants. Its enzyme activity was calculated >30U/mg using the absorbance A275 as the vertical axis and different concentrations of a tyrosine as the horizontal axis. The working concentration of proteinase K, usually determined by the temperature, PH, and whether the buffer used contains SDS and urea, is 20mg/mL. This recombinant proteinase K protein is in-stock now. Proteinase K has a wide range of applications. In nucleic acid extraction and isolation, proteinase K can be used to degrade Dnase/Rnase and DNA-bound histones and lyse cells or tissues. It also digests protein impurities especially endotoxin and separates target nucleic acids from proteins in biological samples, thus improving the purity and safety of nucleic acids and production efficiency. The nucleic acids isolated by proteinase K is often used for the amplification reaction. Denaturing agents such as SDS (1%) can increase the enzymatic activity of proteinase K. In in-situ hybridization, it removes the proteins around the target DNA to facilitate probe infiltration and improves detection sensitivity. Proteinase K is also used to analyze the membrane structure through the modification of proteins on the cell surface. The nucleic acid-based real-time reverse transcriptase-polymerase chain reaction (RT-PCR) test is the primary tool to diagnose the SRAS-CoV-2 infection. Proteinase K is often used to remove unwanted proteins present in the nasal/pharyngeal swab samples and inactivate nucleases that could degrade DNA or RNA during isolation and purification procedures, thus increasing the yield of extracted RNA and improving the detection of viral RNA by RT-PCR. |
Purity | Greater than 95% as determined by SDS-PAGE. |
Activity | Using the absorbance A275 as the vertical axis and different concentrations of tyrosine as the horizontal axis, a standard curve was drawn, and the enzyme activity was calculated >30U/mg. |
Target Names | PROK |
Uniprot No. | P06873 |
Alternative Names | Endopeptidase K,Tritirachium alkaline proteinase |
Species | Tritirachium album (Engyodontium album) |
Mol. Weight | 28.9 kDa |
Form |
Lyophilized powder Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand. |
Buffer | If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.
Note: If you have any special requirement for the glycerol content, please remark when you place the order. If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0. |
Reconstitution | We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference. |
Troubleshooting and FAQs |
Protein FAQs |
Storage Condition | Store at -20°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles. |
Shelf Life | The shelf life of the lyophilized form is 24 months at -20°C/-80°C, 18 months at 4°C, and 12 months at room temperature. |
Lead Time | Basically, we can dispatch the products out in 3-7 working days after receiving your orders. Delivery time may differ from different purchasing way or location, please kindly consult your local distributors for specific delivery time. |
Notes | Repeated freezing and thawing is not recommended. |
Datasheet & COA | Please contact us to get it. |
Applications : Antigen, Drug related studies, Protein structure analysis (Crystal / electron microscope)
Review: Used proteinase K to improve the purity of the extracted DNA from a Bacillus thermoamylovorus strain producing a thermotolerant lipase by digesting contaminating cell proteins. To check the effectiveness of proteinase K to inhibit the activity of purified lipase in the absence and presence of PMSF.
By Shamsher S Kanwar
What is the activity of Proteinase K in common buffers?
What is the recommended pH for Proteinase K to keep its enzyme activity?
What is the main applications of Proteinase K?
How to dissolve proteinase K powder? What should I do if the dissolution is not complete or the dissolution time is longer ?
What is the long-term storage temperature of proteinase K from CUSABIO?
Whether the form of proteinase K is liquid or powder, will repeated freezing and thawing affect the activity?
What is the recommended working concentration of Proteinase K from CUSABIO?
What is the optimal temperature for proteinase K activation? And how to inactivate proteinase K?
Proteinase K digest protein and remove contamination, but it doesn’t digest itself. Why?
How long do the samples need incubating with Proteinase K?
Does EDTA inactivate proteinase K?
How does Proteinase K works in DNA extraction?
Can proteinase K degrade DNA?
Instead of degrading the DNA, protease K digests the proteins surrounding the DNA, allowing it to be isolated intact.
What does proteinase K do to the cell?
Proteinase K can lyse or break down the cell membrane and release the intercellular materials including DNA and RNA.
How do you use proteinase K?
Dissolve lyophilized proteinase K in deionized sterile water. We recommend to add 10 mM Tris HCl, pH 7.5, 20 mM calcium chloride, and 50% glycerol, and then aliquot for long-term storage at -20/-80°C.
Why do we denature proteinase K?
Because proteinase K may degrade the useful proteins in the downstream applications of isolated nucleic acids.
How do you make proteinase K buffer?
The best buffer for proteinase K will be different in various applications. Proteinase K is active over a wide pH range (4-12.5) and relatively high temperature (optimum temperature 50-65 ˚C). Generally, proteinase K is stable and keeps highly active in buffers that contain denaturing reagents such as urea, sodium dodecyl sulfate (SDS), and guanidinium salts.
Why do we add proteinase K to our DNA extractions?
Proteinase K is used to lyse cells, disrupt tissue integrity, and degrade various proteins to make the pure DNA is fully released during DNA extraction. It also inactivates DNase present in the sample, ensuring DNA whole and intact. The wide temperature range and relatively strong pH tolerance of proteinase K also contribute to its extensive availability in nucleic acid extraction.
Function |
Hydrolyzes keratin at aromatic and hydrophobic residues.
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Protein Families | Peptidase S8 family |