Cat. No. / ID: EN31-005
dsDNase HL (500 U) is a 43.3 kDa heat-labile recombinant endonuclease derived from a cold water eukaryotic organism expressed in Pichia pastoris. The enzyme displays high specific activity towards double-stranded DNA, leaving single-stranded DNA or RNA undamaged in standard conditions.
It is supplied with 20 mM Tris-HCl, pH 8.0; 20 mM NaCl; 2 mM MgCl2 ; 1 mM CaCl2 ; 50% (v/v) glycerol.
One unit is defined as the amount of enzyme that causes an increase in absorbance at 260 nm of 1.0 in 30 minutes at 37°C in 50 mM Tris-HCl buffer, pH 8.0 (25°C) supplemented with 5 mM MgCl2, 0.1 mg/mL BSA and 0.5 mg/mL herring sperm DNA as a substrate.
|RNase activity||None detected|
|Proteolytic activity||None detected|
dsDNase HL can be quickly inactivated by heat treatment in moderate temperatures. It is intended for applications where the presence of dsDNA influences experiments’ results in thermo-sensitive applications. It is also beneficial for the rapid and safe purification of RNA or protein samples from contaminating DNA.
The enzyme hydrolyzes phosphodiester linkages yielding oligonucleotides with a 5′-phosphate and 3′-hydroxyl groups.
Protein concentration was determined by densitometry of SDS-PAGE electrophoresis using Coomassie Blue detection assay.
The presence of RNase activity was determined after incubation of 5 U dsDNase HL with 1 μg of total eukaryotic RNA in a 20 μL volume for 1 hour at 37°C.
The presence of proteolytic activity was determined by SDS-PAGE with Coomassie Blue detection following incubation of 1 μg of BSA with 100 U of enzyme for 20 hours at 37°C.
A 1.575 mL reaction volume containing herring sperm DNA as a substrate is incubated for 30 minutes at pH 8 at 37°C with an enzyme that degrades DNA to acid-soluble oligonucleotides. One unit of the enzyme causes an increase in absorbance at A260 of 1.0.
This is used for applications such as: