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導入
Proteinase K is a stable serine protease with broad substrate specificity. It degrades many proteins in the native state even in the presence of detergents. It is not activated by metalions, キレート剤 (例えば, EDTA), スルフヒドリル試薬, or by tripsinor chymotrypsin inhibitors. It is stable over a wide pH range (4-12.5), pH 6.5 ～ 9.5 で最適な活性を発揮. Activity can be stimulated by addition of denaturing agents (SDS と尿素). Rapid denaturation of the enzyme occurs at temperature above 70°C. Autolysis of the enzyme occurs increasingly at alkaline pH. しかし, Proteinase K is not completely inactivated by autolysis. Some enzyme fragments continue to maintain their complete proteolytic activity, even after extensive autolysis.
Proteinase K is frequently used in molecular biology applications to digest unwanted proteins, such as nucleases in DNA or RNA preparations from microorganisms, cultured cells, and plants. The enzyme is typically used at 50–200 μg/ml in nucleic acid preparations at pH 7.5-8.0 および37〜55℃. インキュベーション時間は次のように異なります。 30 分まで 18 時間.
詳細
仕様
| 特徴 | 仕様 |
| 分子量 | 29.3kDa |
| 等電点 | 8.9 |
| 純度 | 95% (SDS-PAGE分析) |
| 特定の活動 | ≥600 Units/ml |
| 温度特性 | 70℃ (maximum activity temperature) (effective activity temperature is 37-70°C) |
| PH特性 | 4.0-12.0, 最適な範囲はph7.5〜11.5です。. Indigestive juice, 65 degrees |
| 保存条件 | It is recommended to store at low temperature to ensure the stability of activity to the greatest extent (常温で輸送または保管しても酵素活性は低下しません). The shelf life at room temperature is up to 1year. |
| 利用方法 | Proteinase K was added to the digestive solutionor lysate until the final concentration was 50-200ug/ml and incubated at 55~70°C. 反応後のプロテイナーゼKは磁気ビーズ法により除去・不活化可能, over column method or phenol chloroform extraction. プロテアーゼ K は、95°C でのインキュベーションによって不活化できます。 3 分または70℃ 15 分 |
| 核酸残基の検出 | Qubit quantification, ≤1ppm. No human source DNA contamination (not detected by real time PCR) |
| ヌクレアーゼの検出 | DNase が検出されない RNase が検出されない |
| 推奨アプリケーション | Tissue and cell DNA extraction, Viral Nucleic acid Isolation |
利点
- Ready to use normal temperature liquid protease K
- Convenient to transport and store at room temperature
- High activity, 20 mg/ml, >600 mAu/ml
注文情報
| コンテンツ | C12103 | C12104 |
| プロテイナーゼ K ソリューション (20mg/ml) | 10 ミリリットル | 1000 ミリリットル |
レビュー
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