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UCF.ME® UltraNuclease GMP-grade

Product SKU

Specifications

Price

20157ES25

25 KU

$173.70

20157ES50

50 KU

$303.40

20157ES60

100 KU

$501.03

20157ES80

1MU(1000KU)

20157ES90

5MU(5000KU)

Product Description

UltraNuclease is a nonspecific endonuclease recombinant from Serratia Marcescen and expressed in Escherichia coli (E. coli). The sequence of the enzyme is consistent with Benzonase (Merck) It catalyzes nonspecific endonucleolytic cleavage of DNA and RNA of all formats (double-stranded, single-stranded, linear, circular, naturally occurring, or denatured) to yield 5'-monophosphate oligonucleotides of 2-5 bp under a broad range of conditions (6 M Urea,0.1 M Guanidine HCl,0.4% Triton X-100,0.1% SDS,1 mM EDTA,1 mM PMSF). This product has been widely used for nucleic acid digestion for numerous applications.

The product is engineered, expressed, purified, and manufactured under the GMP environments. It is often used to reduce cell supernatant or cell lysate viscosity, thus increasing protein purification efficiency and enhancing protein functional. The product can also be used to reduce host nucleic acid residues to pg-grade, improving the performance and safety of biology products of applications including virus purification, vaccine manufacturing, and protein/polysaccharide pharmaceutical manufacturing. Besides, the product can also be applied to prevent clumping of human peripheral blood mononuclear cells (PBMC) in cell therapy and vaccine development.

The product is provided in the form of a sterilized reagent, eluted in buffer (20 mM Tris-HCl pH 8.0, 2 mM MgCl2, 20 mM NaCl, 50% glycerin), with the appearance of colorless, transparent liquid.

Product Properties

Expression HostEscherichia coli (E. coli)
SynonymBenzonase; Endonuclease; Nonspecific Endonuclease
Molecular Weight26.5 kDa
Isoelectric point6.85
Purity≥ 99%(SDS-PAGE)
Enzyme Activity250-300 U/μL
Specific Activity≥1.5×106 U/mg protein
Optimum pH8.0(working pH range: 6-10)
Optimum Temperature37℃(working temperature range: 0-42℃)
Cofactor1-10 mM Mg2+
ProteaseNegative
MicroorganismNegative
Endotoxin < 0.25 EU/1000U
Host protein < 10 ppm(μg/mL)
Storage Buffer20 mM Tris-HCl pH8.0,2 mM MgCl2,20 mM NaCl,50% glycerin
Unit DefinitionThe definition of one Activity unit (U) is the amount of enzyme used to change the absorption value of △A260 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).

Shipping and Storage

The product is shipped with ice pack and can be stored at -20℃ for 1 year. If the product is opened and has been stored at 4℃ for more than a week, we recommend filtering the product to prevent microorganism contamination.

Product Note

For your safety and health, please wear lab coats and disposable gloves for operation.

Recommended conditions

Parameter

Optimal Condition

Effective Condition

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 Ion Concentration

0-10 mM

0-100 mM

Recommended Reaction Time (37℃, 2 mM Mg2+, pH 8.0)

UltraNuclease Amount (Final Concentration)

Treatment Time

0.25 U/mL

> 10 h

2.5 U/mL

> 4 h

25 U/mL

30 min

Protocol

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.

Suspension cells: collect the cells by centrifugation, wash the cells with PBS, centrifuge at 6,000 rpm for 10 min, 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 (1 g of cell pellet contains about 109 cells).

3. Enzyme Treatment

1)Add 1-5 mM MgCl2 to the reaction system, and adjust the pH to 8-9.

2)Add UltraNuclease according to the ratio of 250 Units to digest 1 g of cell pellets, incubate at 37°C for more than 30 minutes. Please refer to the “Recommended Reaction Time” form above 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.

15221439726

15221439726