Ultra Nuclease GMP-grade
Cat No: HC2016A
UltraNuclease GMP-grade is expressed and purified in Escherichia coli (E.coli) by genetically engineered and prepared under the GMP environments. It can reduce the viscosity of cell supernatant and cell lysate in scientific research, increase protein purification efficiency and enhance protein functional research. The product can also reduce host nucleic acid residues to pg-grade, improving the performance and safety of biological products of applications including virus purification, vaccine manufacturing, and protein/polysaccharide pharmaceutical manufacturing. Besides, the product can also be applied to prevent the clumping of human peripheral blood mononuclear cells (PBMC) in cell therapy and vaccine development.
UltraNuclease is provided in the form of a sterilized reagent, eluted in buffer (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), with the appearance of a colorless, transparent liquid. This product is produced by GMP process requirements and provided in a liquid form.
Components
UltraNuclease GMP-grade (250 U/μL)
Storage Conditions
The product is shipped with dry ice and can be stored at -25℃~-15°C for two years.
If the product is opened and has been stored at 4℃ for more than a week, we recommend filtering the product to prevent microbial contamination.
Specifications
|
Expression Host |
Recombinant E. coli with UltraNuclease gene |
|
Molecular Weight |
26.5 kDa |
|
soelectric point |
6.85 |
|
Purity |
≥99% (SDS-PAGE) |
|
Storage Buffer |
20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Unit Definition |
The definition of one Activity unit (U) is the amount of enzyme used to change the absorption value of ΔA260 by 1.0 in 30 minutes in a 2. 625 mL reaction system at 37℃ with a pH of 8.0 (equivalent to complete digestion of 37 μg salmon sperm DNA into oligonucleotides). |
Instructions
1. Sample Collection
Adherent cells: remove the medium, wash the cells with PBS, and remove the supernatant.
Suspension cells: collect the cells by centrifugation, wash the cells with PBS, centrifuge at 6,000 rpm for 10 min, collect the pellet.
Escherichia coli: collect the bacteria by centrifugation, wash once with PBS, centrifuge at 8,000 rpm for 5 min, and collect the pellet.
2. Sample Treatment
Treat the collected cell pellets with lysis buffer at the ratio of mass(g) to volume(mL)1:(10-20), or by mechanical or chemical methods on ice or at room temperature (1g of cell pellet contains about
109 cells).
3. Enzyme Treatment
Add 1-5mM MgCl to the reaction system and adjust the pH to 8-9.
Add UltraNuclease according to the ratio of 250 Units to digest 1 g of cell pellets, incubate at 37℃ for more than 30 minutes. Please refer to the “Recommended Reaction Time” form to choose the duration of the treatment.
4. Supernatant Collection
Centrifuge at 12,000 rpm for 30 minutes and collect the supernatant.
Note: If the solution is acidic or alkaline, or contains high concentrations of salt, detergents, or denaturants, please increase the enzyme dosage or extend the treatment time accordingly.
Recommended reaction conditions
|
Parameter |
Optimal Condition |
Effective Conditior |
|
Mg²+Concentration |
1-5 mM |
1-10 mM |
|
pH |
8-9 |
6-10 |
|
Temperature |
37℃ |
0-42℃ |
|
DTT Concentration |
0-100 mM |
>0 mM |
|
Mercaptoethanol Concentration |
0-100 mM |
>0 mM |
|
Monovalent Cation Concentration |
0-20 mM |
0-150 mM |
|
Phosphate lon Concentration |
0-10 mM |
0-100 mM |
Recommended Reaction Time (37℃, 2 mM Mg²+, pH 8.0)
|
UltraNuclease Amount (Final Concentration) |
Reaction Time |
|
0.25 U/mL |
>10 h |
|
2.5 U/mL |
>4h |
|
25 U/mL |
30 min |
Notes:
Please wear the necessary PPE, such lab coat and gloves, to ensure your health and safety!
