|Manufactured By:||Sino Biological Inc.|
|Synonym:||SuperNuclease, Nuclease, Serratia marcescens’ extracellular endonuclease|
|Protein Construction:||SuperNuclease is a recombined Serratia marcescens’ extracellular endonuclease. The sequence is same with Benzonase® (Benzonase is a trademark of Merck KGaA, Darmstadt, Germany).|
|Expression Host:||Escherichia coli (E. coli).|
|Purity:||> 95 % as determined by SDS-PAGE.
> 99 % as determined by SEC-HPLC Analysis.
|Endotoxin:||< 0.01 EU/1000 units as determined by the LAL method.|
|Activity:||Measured by its ability to cleave Salmon sperm DNA substrate (Sigma, Catalog # D1626). One unit is defined as the amount of enzyme that causes a ΔA260 of 1.0 in 30 minutes at 37°C.
The specific activity is ≥ 1.1×106 units/mg.
|Predicted N terminal:||Three isoforms with different N terminal may be found from the compound—Sm1 (22D-266N), Sm2 (23T-266N) and Sm3 (25E-266N), the activity analysis shows that they were functionally equivalent.|
|Molecular Mass:||The SuperNuclease comprises 266 amino acids and has a calculated molecular mass of Sm1 (26708.2 Da), Sm2 (26591.8 Da) and Sm3 (26376.4 Da). The apparent molecular mass of SuperNuclease is approximately 26.5 Kda.|
|Storage:||Store it under sterile conditions at -20℃ to -80℃ upon receiving for at least 12 months. Recommend to aliquot the protein into smaller quantities for optimal storage. Avoid repeated freeze-thaw cycles.|
1). Lysis buffer preparation:
The lysis buffer should be suitable for the protein as well as the downstream purification processes, and so on;
2). Resuspend cell plates in lysis buffer:
The ratio of lysis buffer (mL) against the gram of cell can be (10-20):1;
3). Add SuperNuclease (250 units to 1 g cells):
It is well recommended to optimize the amount of SuperNuclease;
4). Lysate cell by mechanical or chemical methods on ice or at room temperature.
Methods: Ultrasonic disruption, High Pressure Homogenizer, tissue homogenizer, and so on;
5). Obtain clear cell lysate supernatant by centrifugation at ~12,000 rpm for 0.5 hour.