ProNuclease - 汇融科技

E.Coli Recombinant nuclease

BETAGENE ProNuclease

Specification

CAS号	EC号	MDL编号	货号
9025-65-4	3.1.30.2(BRENDA，IUBMB)	MFCD00131010	GA01.01

Specification Sheet

MSDS

COA

ProNuclease by BETAGENE

Ultimate solution for nucleic acid residue

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.

Unit

158997890645513199695db917eed1868815883f56b63

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.

Application

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.

Buffer

Storage buffer: 20 mM Tris-HCl （pH 8.0），2 mM MgCl₂，20 mM NaCl，and 50% Glycerol

Cautions

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.

QA

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)

Safety data

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

Specs

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℃

High activity
Complete removal
Multiple applications
High tolerance

Specs
FAQs

Composition

E. Coli Recombinant Nuclease

Cat. No.

GA01.01/02/03/04

Clear, Colorless

Appearance

Host

E.coli

Molecule Weight

About 30 kD

AA Sequence

ADTLESIDNCAVGCPTGGSSNVSIVRHAYTLNNNSTTKFANWVAYHITKDTPASGKTRNWKTDPALNP

ADTLAPADYTGANAALKVDRGHQAPLASLAGVSDWESLNYLSNITPQKSDLNQGAWARLEDQERKLI

DRADISSVYTVTGPLYERDMGKLPGTQKAHTIPSAYWKVIFINNSPAVNHYAAFLFDQNTPKGADFCQF

RVTVDEIEKRTGLIIWAGLPDDVQASLKSKPGVLPELMGCKN

Acitivity

≥ 250kU/mL，DNA；pH 8.0；30min；37℃

Store Buffer

20 mM Tris-HCl（pH 8.0）, 2 mM MgCl2, 20 mM NaCl, and 50% glycerol

Purity

> 99 % by SDS-PAGE analyses. > 99 % by HPLC analyses.

Usage

This GMP product can be for research use or further manufacturing use.

-20℃ over one year

Storage

Protease

Negative

Endotoxines

< 0.01 EU/kU

Spec. activity

1.5 x 10⁶ kU/mL；calculated on protein

Different application steps

Virus purification: usually after virus is clarified;

E. coli recombinant protein purification: applied when the cells are lysed;

Cell lysate clumping: added directly to the culture medium for co-cultivation.

What to do if the reaction temperature cannot reach 37 ℃?

Enzyme activity is affected by temperature, treatment time and enzyme activity unit. If the temperature cannot reach 37 ℃, the enzyme dosage can be increased or the incubation time can be prolonged.

What are the optimal reaction conditions for ProNuclease?

The optimal reaction conditions for the enzyme are pH 8.0-9.2 and 37 ° C for half an hour. Beyond this range, the enzyme is still active, but the reaction time needs to be extended.

Is ProNuclease compatible with protease inhibitors?

Yes, but it should be noted that many protease inhibitors contain EDTA;

When EDTA levels start to exceed 1 mM, EDTA will inhibit some nuclease activity.

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