| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 500mg |
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| Other Sizes |
| Targets |
Proteins (broad substrate specificity). Proteinase K is a serine protease that preferentially cleaves the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids (hydrophobic residues).
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| ln Vitro |
The amid link between MTX and BSA as well as the amidic bond within the BSA structure can be broken by the enzyme proteinase K[2]. Guidelines (This is our suggested protocol; it should be adjusted to meet your unique needs; it is merely meant to serve as a guide). Without the need for RNA extraction, the SARS-COV2 viral component can be directly detected from inoculation viral transport medium (VTM) using proteinase K, which has better protein-degrading capabilities[3]. The steps involved in the rRTPCR detection protocol for SARS-CoV2 are as follows: 1. Divide the VTM of each SARS-COV2 sample into three 45 μL EP tubes; 2. Add 2.5 µL (20 mg/mL) concentrated proteinase K (PK), resulting in a final concentration of PK ~1 mg/ml VTM; 3. Incubate at 25 °C for 5 min; 4. Incubate at 60 °C for 15 min; 5. Incubate at 98 °C for 10 min, and store the samples at 4 °C. 6. Use 5 µL of extracted RNA as template to carry out the rRTPCR assay; 7. Utilize the BIORAD CFX96™ Real-Time Thermocycler for rRTPCR detection, with a positive standard of threshold cycle (Ct) values less than 40. Proteinase K raises the concentration of nucleic acids recovered from sputum samples and enhances their homogenization effects and positive rates[4]. The procedure for extracting influenza A virus (IAV) nucleic acid is as follows: 1. Divide the specimen into three 500 μL EP tubes; 2. Add 500 μL of Proteinase K buffer (0.4 mg/mL); 3. Incubate at 55 °C for 25 minutes, vortexing every five minutes; 4. Extract RNA using the Automated Nucleic Acid Extraction System (Zhijiang Biotechnology Co., Ltd., Shanghai, China) using 200 μL samples; 5. Assess the A260/A280 ratio of pure RNA products, using a standard range of 1.8 to 2.0.
Proteinase K has broad substrate specificity and high proteolytic activity. It effectively degrades many native proteins even in the presence of detergents such as 1% Triton X-100 or 0.5% SDS. It is active over a wide pH range (4-12) and temperature range (0-75degC), making it valuable for diverse biochemical applications. |
| ln Vivo |
Proteinase K is not used as a systemic therapeutic agent. However, it has been studied for its ability to degrade misfolded or aggregated proteins in localized settings. In animal studies, it has been used ex vivo for tissue digestion and in vivo for investigating protease inhibitor activity, though its systemic use is limited due to immunogenicity.
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| Enzyme Assay |
For enzyme activity assays, Proteinase K is incubated with a chromogenic substrate such as succinyl-Ala-Ala-Pro-Phe-p-nitroaniline (s-AAPF-pNA) in assay buffer (50 mM Tris-HCl pH 7.5, 5 mM CaCl2). Hydrolysis of the substrate releases p-nitroaniline, which is quantified by measuring absorbance at 405 nm. One unit is defined as the amount of enzyme that hydrolyzes 1 micromol of substrate per minute.
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| Cell Assay |
Proteinase K is used in cell culture for harvesting adherent cells by gentle proteolytic detachment from culture vessels, alternative to trypsin. It can also be used to digest cellular proteins in lysates prior to DNA or RNA extraction. For cell viability assays, it is used in the preparation of cell lysates for downstream applications such as protein degradation studies.
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| Animal Protocol |
For in vivo inhibitor studies, animals are injected with a putative Proteinase K inhibitor or control. Tissues are harvested, and extracts are incubated with Proteinase K in the presence of a chromogenic substrate (s-AAPF-pNA). Inhibition is determined by comparing protease activity to untreated controls. This ex vivo approach assesses systemic protease inhibitor activity.
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| ADME/Pharmacokinetics |
Proteinase K is not used as a drug, so its human pharmacokinetics are not relevant. As an enzyme, it degrades proteins rapidly in vivo and has a short biological half-life. For in vitro use, it is stable at 4degC for short-term storage and at -20degC for long-term storage.
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| Toxicity/Toxicokinetics |
Proteinase K is generally considered safe when handled with standard laboratory precautions. In its purified form, it is not known to be acutely toxic. However, as an active protease, it can cause tissue damage if injected at high concentrations. It is intended for research use only.
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| References | |
| Additional Infomation |
Proteinase K (CAS: 39450-01-6) has a molecular weight of approximately 28.9 kDa. It is often supplied as a lyophilized powder with an activity >30 units/mg protein. It is also commercially available as a recombinant protein expressed in Pichia pastoris. It is a standard reagent in molecular biology laboratories.
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| Molecular Formula |
C29H27N2O12P
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|---|---|
| Molecular Weight |
626.5046
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| Exact Mass |
391.2
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| CAS # |
39450-01-6
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| Appearance |
White to off-white solid powder
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| Density |
1.3±0.1 g/cm3
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| Index of Refraction |
1.648
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| LogP |
2.5
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O: 25 mg/mL
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| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (Infinity mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5962 mL | 7.9808 mL | 15.9617 mL | |
| 5 mM | 0.3192 mL | 1.5962 mL | 3.1923 mL | |
| 10 mM | 0.1596 mL | 0.7981 mL | 1.5962 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
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