SuperNuclease

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General Information

Manufactured By: Sino Biological Inc.
Synonym: Nuclease, Serratia marcescens’ extracellular endonuclease
Protein Construction: SuperNuclease is a recombined Serratia marcescens’ extracellular endonuclease.
Source: Aequorea victoria
Expression Host: Escherichia coli (E. coli).

QC Testing

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.
Stability: Samples are stable for up to twelve months from date of receipt at -20℃ to -80℃.
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.

SDS-PAGE:

SEC-HPLC:

Usage Guide

Storage: Store it under sterile conditions at -20℃ to -80℃ upon receiving. Recommend to aliquot the protein into smaller quantities for optimal storage.

Protocol

Large scale cell lysis treatment:

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.

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