CAS号 | EC号 | MDL编号 | 货号 |
9025-65-4 | 3.1.30.2(BRENDA，IUBMB) | MFCD00131010 | GA01.01 |
Introduction
BETAGENE® ProNuclease is a non-specific endonuclease. It hydrolyzes internal phosphodiester bonds present between the nucleotides, all free nucleic acids present in solution are reduced to 5'-monophosphate terminated oligonucleotides which are 3 to 8 bases in length. ProNuclease is able to degrade different types of DNA and RNA, whether single-stranded, double-stranded, linear, circular or supercoiled. ProNuclease is widely used in biological proteins purification.
BG-Pronuclease is a genetically engineered endonuclease produced in E. coli. The molecular weight is about 30 kDa each. The Purity is over 99 %（SDS-PAGE、HPLC），the minimum specific activity 1.5 x 106 units/mg, the enzyme activity is over 250U/μL，and the total endotoxin level is below 0.01 EU/kU.
Features
Removing exogenous nucleic acids from vaccin and genetic medicine to improve product safety;
Reducing the viscosity of solution caused by nucleic acids, shortening the processing time, and improving the efficiency of subsequent purification process and the final product yield;
Removing nucleic acids entangled on the surface of various particles, which is conducive to full release and purification of target protein;
Improving the recovery of soluble inclusion bodies after renaturation;
Improving the sample discernibility and recovery rate of ELISA, column chromatography, two-dimensional electrophoresis and Western blot analysis.
Removal of ProNuclease residue
Just like any other miscellaneous protein, ProNuclease in bacterial or cell lysate can be removed during purification. ProNuclease in virus or vaccine, however, can be removed by gel filtration such as Sepharose 6FF because of the difference in molecular weights. You can test the residual amount of nuclease with commercial kit.
One unit (U) of ProNuclease is defined as the amount of enzyme that causes a change of 1.0 in A260 within 30 minutes, at 37 ℃, pH 8.0, 50mM Tris-HCl, 1mM MgCl2, 100 μg/mL BSA, and 1mg/mL sonicated salmon sperm DNA.
To reduce the viscosity of cell supernatants and cell lysates, increasing protein purification efficiency, and to effectively prevent clumping of human peripheral blood mononuclear cells (PBMC) in cell therapy and vaccine research. It is widely used to remove nucleic acids from biological products.
Storage buffer: 20 mM Tris-HCl （pH 8.0），2 mM MgCl2，20 mM NaCl，and 50% Glycerol
The optimal condition of reaction: 30 mins at pH 8.0 - 9.2, 37 °C. Beyond this range the product will still be active, but the duration of reaction should be appropriately extended;
Magnesium ions are essential to product activity. The recommended concentration of magnesium ion is 2 mM. Metal chelating agent such as EDTA needs to be removed if existed in the sample, or excess magnesium ions are required to neutralize EDTA;
The product acitivity is relatively stable. Room temperature will not impact the stability in the short term. However store at below -20℃ in the long term. Avoid repeated freeze-thaw.
Quality Assurance
| Items | Specifications | Standard |
Appearence | Colorless, transparent | Visual |
Identity | Positive | Chinese Pharmacopoeia 2020 Edition (3401) |
Host protein residue | ≤ 0.005% | Chinese Pharmacopoeia 2020 Edition (0731) |
Visible particles | No grandules | Chinese Pharmacopoeia 2020 Edition (0904) |
pH | 7.0 - 9.0 | Chinese Pharmacopoeia 2020 Edition (0631) |
Activity | ≥ 250kU/mL | One unit (U) of ProNuclease is defined as the amount of enzyme that causes a change of 1.0 in A260 within 30 minutes, at 37 ℃, pH 8.0, 50mM Tris-HCl, 1mM MgCl2, 100 μg/mL BSA, and 1mg/mL sonicated salmon sperm DNA. |
Purity | ≥ 99% | Chinese Pharmacopoeia 2020 Edition (0541); SEC-HPLC |
Protease | Negative | 1 kU/ml, incubate with substrate at 37℃ for 60 min |
Endotoxin | ≤ 0.01 EU/kU | Chinese Pharmacopoeia 2020 Edition (1143) |
Sterility test | Negative | Chinese Pharmacopoeia 2020 Edition (1101) |
Pathogen | Negative | Chinese Pharmacopoeia 2020 Edition (3306) |
Mycoplasma | Negative | Mycoplasma test kit; PCR |
Heavy metal residue | ≤ 10 ppm | Chinese Pharmacopoeia 2020 Edition (0821) |
Appearence：Colorless |
Visual |
Identity：Positive |
Chinese Pharmacopoeia 2020 Edition (3401) |
Host protein residue ≤ 0.005% |
Chinese Pharmacopoeia 2020 Edition (0731) |
pH ：7.0 - 9.0 |
Chinese Pharmacopoeia 2020 Edition (0631) |
Activity ≥ 250kU/mL |
One unit (U) of ProNuclease is defined as the amount of enzyme that causes a change of 1.0 in A260 within 30 minutes, at 37 ℃, pH 8.0, 50mM Tris-HCl, 1mM MgCl2, 100 μg/mL BSA, and 1mg/mL sonicated salmon sperm DNA. |
Purity ≥ 99% |
Chinese Pharmacopoeia 2020 Edition (0541)、 SEC-HPLC |
Protease：Negative |
1 kU/mL, incubate with substrate at 37 °C for 60 min |
Endotoxines ≤ 0.01 EU/kU |
Chinese Pharmacopoeia 2020 Edition (1143) |
Sterility test：Negative |
Chinese Pharmacopoeia 2020 Edition (1101) |
Pathogen：Negative |
Chinese Pharmacopoeia 2020 Edition (3306) |
Mycoplasma：Negative |
Mycoplasma test kit; PCR |
Heavy metal residue ≤ 10 ppm |
Chinese Pharmacopoeia 2020 Edition (0821) |
储存分类代码 | WGK | 闪电（F) |
10-Combustible liquids | WGK 1 | Not Applicable |
闪电（C) | 个人防护装备 | |
Not Applicable | Eye shields, gloves | |
COMPOSITION E.COLI RECOMBINANT NUCLEASE | |
MOLECULE WEIGHT APPROX. 30kD | |
CAT.No GA01.01/02/03/04 | APPEARENCE COLORLESS |
ENDOTOXINES < 0.01 EU/kU | PROTEASE NEGATIVE |
PURITY > 95 % BY SDS-PAGE. > 95 % BY HPLC | |
ACTIVITY ≥ 250 kU/mL，DNA; pH 8.0；30MIN; 37℃ | |
SPECS. ACTIVITY 1.5 X 106 kU/mL；CALCULATED ON PROTEIN | |
AA SEQUENCE ADTLESIDNCAVGCPTGGSSNVSIVRHAYTLNNNSTTKFANWVAYHITKDTPASGKTRNWKTDPALNPADTLAPADYTGANAALKVDRGHQAPLASLAGVSDWESLNYSNITPQKSDLNQGAWARLEDQERKLIDRADISSVYTVTGPLYERDMGKLPGTQKAHTIPSAYWKVIFINNSPAVNHYAAFLFDQNTPKGADFCQFRVTVDEIEKRTGLIIWAGLPDDVQASLKSKPGVLPELMGCKN | |
FORMULATION 20 mM TRIS-HCl (pH 8.0)，2 mM MgCl2，20 mM NaCl，50% GLYCEROL | |
USAGE FOR REASEARCH OR FURTHER MANUFACURE USE | |
STORAGE -20℃ | |
Specs
FAQs
1.5 x 106 kU/mL；calculated on protein
