介紹
- Proteinase K Characteristics:
- Stable serine protease with broad substrate specificity.
- Degrades many proteins in the native state even in the presence of detergents.
- Not activated by metal ions, 螯合劑 (例如, 乙二胺四乙酸), 巰基試劑, or trypsin or chymotrypsin inhibitors.
- Stable over a wide pH range (4-12.5), 在 pH 6.5–9.5 時具有最佳活性.
- Activity can be stimulated by denaturing agents (例如, 安全資料表 和尿素).
- Rapid denaturation occurs at temperatures above 70°C.
- Autolysis increases at alkaline pH, but some enzyme fragments maintain complete proteolytic activity even after extensive autolysis.
- 應用領域:
- Frequently used in molecular biology to digest unwanted proteins, such as nucleases in DNA or RNA preparations.
- Typically used at 50–200 μg/ml in nucleic acid preparations.
- Optimal conditions: 酸鹼度 7.5-8.0 和37-55°C.
- 孵化時間各不相同 30 分鐘到 18 小時.
規格
| 特徵 | 規格 |
| 分子量 | 29.3kDa |
| 等電點 | 8.9 |
| 外貌 | White lyophilized powder |
| 純度 | 95% (SDS-PAGE分析) |
| 具體活性 | ≥34 Units/mg protein |
| 溫度特性 | The effective activity temperature is 37-70°C, and the enzyme activity at 65°C is twice that at 25°C. |
| PH特性 | 4.0-12.0, 最佳範圍為ph7.5-11.5 |
| 保存條件 | It is recommended to store at -20°C to ensure its stability to the greatest extent (常溫運輸或儲存不會降低酵素活性). The shelf life is up to 3 years at -20°C and 2 years at 2~8°C. |
| 使用方法 | Prepare 20mg/ml with the solution, add proteinase K to the digestive solution or lysate until the final concentration is 50-200ug/ml, and incubate at 55~70°C. 反應後的蛋白酶K可透過磁珠法去除或滅活, column method or phenol-chloroform extraction. 95℃ 孵育可滅活蛋白酶 K 3 分鐘或 70°C 15 分分鐘. |
| 核酸殘留檢測 | Qubit did not detect Human DNA contamination not detected (即時的 聚合酶鍊式反應) Bacterial DNA contamination not detected (16S universal primer PCR, 30 週期) Fungal DNA contamination not detected (ITS Primer PCR) |
| 核酸酶檢測 | 未檢測到 DNA 酶 未檢測到 RNase Nickase not detected |
| 推薦應用 | 核酸萃取, circulating DNA extraction, virus nucleic acid extraction |
優點
- Not including RNase, 脫氧核糖核酸酶, and Nickase
- No nucleic acid residue (not detected by Qubit after extracting the whole 100mg protein K)
- No human-source DNA contamination (not detected by Q-PCR)
- No bacterial DNA contamination (not detected by 16S universal primer)
- No fungal DNA contamination (not detected by 1TS universal primer)
- Can be transported and valid for two years at normal temperature in a dry environment.
- Can be used for highly sensitive free DNA extraction, virus nucleic acid extraction, whole blood DNA extraction, ETC.
訂購資訊
| 內容 | C12100 | C12101 | C12102 | PDB-1000 |
| 蛋白酶K, 冷凍乾燥, >30 units/mg of protein | 1 G | 10 G | 100 G | |
| Buffer PDB (20毫摩爾三羥甲基氨基甲烷, 酸鹼度 7.5, 10毫摩爾氯化鈣2, 50% 甘油, 0.1% Preservatives) | 1000 毫升 |
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