Ideal for DNA and RNA clearance
Free of detectable proteolytic activity
Robust over a wide range of operating conditions
Fig.1 The purity of BenzNuclease was determined by SDS-PAGE reduced and staining overnight with Coomassie Blue.
1. Reagents and solutions preparation
50 mM Tris-HCl, 1 mM MgCl2, pH 8.0 ( * In the case of extensive dilution before use, carrier protein such as 0.1 mg/ml HSA or BSA is generally recommended to avoid any enzyme loss from surface adsorption)
1 mg/ml salmon sperm DNA is dissolved overnight at 4 ℃, in reaction buffer, and is then sonicated on ice to obtain a homogenous solution.
Different dilution of nuclease with reaction buffer.
Trichloroacetic acid (TCA)
2. Standard curve establishment
400 μl substrate + 100 μl enzyme of known activity = 500 μl mixture
Incubate the mixture at 37℃ for 30 min.
Stop the reaction by addition of 400 μl cold TCA and incubate on ice for 10 min.
Centrifuge at 8500g for 5 min.
Measure the absorbance of supernatant at 260 nm.
Lot a standard curve with nuclease of known activities for each set of measurements.
3. Measurement of activity
The activity of any unknown nuclease can be determined from a single measurement by means of the standard curve. The specific activity of BenzNuclease is >1.5 x 10e6 unit/mg protein.
Fig 2. Standard curve of nuclease activity for BenzNuclease.
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