Proteinase K蛋白酶K,39450-01-6

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Proteinase K

CAS No. : 39450-01-6

MCE 国际站：Proteinase K

产品活性：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。

研究领域：Metabolic Enzyme/Protease

作用靶点：Ser/Thr Protease

In Vitro: Proteinase K enzyme can be used to break amid bond between MTX and BSA and also amidic bonds in BSA structure.

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.
Protocol of rRTPCR detection of SARS-CoV2:
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™ 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.
Protocol of nucleic acid extraction of influenza A virus (IAV):
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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