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Perkenalan
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, agen pengkelat (for example, EDTA), pereaksi sulfhidril, or by tripsinor chymotrypsin inhibitors. It is stable over a wide pH range (4-12.5), dengan aktivitas optimal pada pH 6,5–9,5. Activity can be stimulated by addition of denaturing agents (SDS dan Urea). Rapid denaturation of the enzyme occurs at temperature above 70°C. Autolysis of the enzyme occurs increasingly at alkaline pH. Namun, 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, sel yang dikultur, and plants. The enzyme is typically used at 50–200 μg/ml in nucleic acid preparations at pH 7.5-8.0 dan 37-55°C. Waktu inkubasi bervariasi dari 30 menit sampai 18 jam.
Detail
Spesifikasi
| Fitur | Spesifikasi |
| Berat molekul | 29.3kDa |
| Titik Isoelektrik | 8.9 |
| Kemurnian | 95% (Analisis SDS-PAGE) |
| Aktivitas tertentu | ≥600 Units/ml |
| Karakteristik suhu | 70°C (maximum activity temperature) (effective activity temperature is 37-70°C) |
| Karakteristik PH | 4.0-12.0, kisaran optimal adalah ph7.5-11.5. Indigestive juice, 65 degrees |
| Kondisi Pelestarian | It is recommended to store at low temperature to ensure the stability of activity to the greatest extent (transportasi atau penyimpanan suhu normal tidak akan mengurangi aktivitas enzim).
The shelf life at room temperature is up to 1year. |
| Metode penggunaan | Proteinase K was added to the digestive solutionor lysate until the final concentration was 50-200ug/ml and incubated at 55~70°C. Proteinase K setelah reaksi dapat dihilangkan atau dinonaktifkan dengan metode manik magnetik, over column method or phenol chloroform extraction. Protease K dapat diinaktivasi dengan inkubasi pada suhu 95°C selama 3 menit atau 70°C selama 15 menit |
| Deteksi residu asam nukleat | Qubit quantification, ≤1ppm.
No human source DNA contamination (not detected by real time PCR) |
| Deteksi nuklease | DNase tidak terdeteksi
RNase tidak terdeteksi |
| Aplikasi yang direkomendasikan | Tissue and cell DNA extraction, Viral Nucleic acid Isolation |
Keuntungan
- Ready to use normal temperature liquid protease K
- Convenient to transport and store at room temperature
- High activity, 20 mg/ml, >600 mAu/ml
Meminta informasi
| Isi | C12103 | C12104 |
| Larutan Proteinase K (20mg/ml) | 10 ml | 1000 ml |
Ulasan
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