Proteinase K (Protease K) 是可用于蛋白消化的一种非特异性丝氨酸蛋白酶。Proteinase K 在有 SDS 或尿素存在的情况下,在很大 pH 值 (4-12)、盐浓度和温度单位内均有活性。Proteinase K 可用于改进提取和检测病毒核酸的方法。
生物活性 | Proteinase K (Protease K) is a nonspecific serine protease that is useful for general digestion of proteins. Proteinase K is active in the presence of SDS or urea and over a wide range of pH (4-12), salt concentrations, and temperatures. Proteinase K can be use for promoting methods of viral nucleic acid extraction, and detection[1][2][3][4]. |
体外研究 (In Vitro) | Proteinase K enzyme can be used to break amid bond between MTX and BSA and also amidic bonds in BSA structure[2].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). Proteinase K with superior protein-degrading property, can be used to detect the SARS-COV2 viral component directly from inoculated viral transport medium (VTM) without RNA extraction[3]. Protocol of rRTPCR detection of SARS-CoV2[3]: 1.Separate VTM of each SARS-COV2 samples into three EP tubes (45 μL per tube); 2.Add 2.5 μL (20 mg/mL concentrated) proteinase K (PK), making a final concentration of PK ~1 mg/ml VTM; 3.Incubate at 25 ℃ for 5 min; 4.Incubate at 60 ℃ for 15 min; 5.Incubate at 98 ℃ for 10 min, and store samples at 4 ℃; 6.Use 5 μL of extracted RNA as template to execute rRTPCR assay; 7.Exert rRTPCR detection by BIORAD CFX96(TM) Real-Time Thermocycler, with a positive standard of threshold cycle (Ct) values less than 40. Proteinase K improves the homogenization effects and positive rates of sputum samples, and increases nucleic acid concentration extracted from it[4]. Protocol of nucleic acid extraction of influenza A virus (IAV)[4]: 1.Separate specimen into three EP tubes (500 μL per tube); 2.Add 500 μL 0.4 mg/mL Proteinase K buffer, vortex for 20 seconds; 3.Incubate at 55 ℃ for 25 min, vortex 5 seconds every 5 min; 4.Extract RNA by using the Automated Nucleic Acid Extraction System through a paramagnetic beads method (Zhijiang Biotechnology Co., Ltd., Shanghai, China) with 200 μL samples; 5.Evaluate A260/A280 ratio of pure RNA products, with a standard ranged from 1.8 to 2.0.
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运输条件 | Room temperature in continental US; may vary elsewhere. |
储存方式 | Powder | -20°C | 3 years | | 4°C | 2 years | In solvent | -80°C | 6 months | | -20°C | 1 month |
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溶解性数据 | In Vitro: H2O : 25 mg/mL(Need ultrasonic) In Vivo: 请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照In Vitro方式配制澄清的储备液,再依次添加助溶剂: ——为保证实验结果的可靠性,澄清的储备液可以根据储存条件,适当保存;体内实验的工作液,建议您现用现配,当天使用;
以下溶剂前显示的百 分比是指该溶剂在您配制终溶液中的体积占比;如在配制过程中出现沉淀、析出现象,可以通过加热和/或超声的方式助溶 1. 请依序添加每种溶剂: PBS Solubility: 50 mg/mL (Infinity mM); Clear solution; Need ultrasonic
*以上所有助溶剂都可在本网站选购。 |