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商品編號


1400010250



品牌


Everest Biotech



公司


Everest Biotech



公司分類(lèi)


Nuclease



Size

250,000 Units




商品信息


Protein Type:
Endonuclease


Source:
E. coli


Unit Definition:
One unit of TurboNuclease converts 1.0 OD
260
of salmon sperm DNA into acid-soluble nucleotides in 30 minutes at 37
o
C in a reaction buffer of 50 mM Tris-HCl, pH 8.0 and 1 mM MgCl
2
. This corresponds to complete digestion of 50 ?g of salmon sperm DNA into oligonucleotides. TurboNuclease has similar specific activity as Benzonase


Storage:
Store TurboNuclease at -20
o
C.


Endotoxin Level:
< 0.1 EU / 1,000 units of TurboNuclease as determined by the Endo-Safe LAL Assay.



Formulation:


250 units/?l in 50 mM Tris-HCl, pH 8.0, 50 mM NaCl, 5 mM MgCl
2
and 50% Glycerol


TurboNuclease Activity and Purity:




50 ?g of salmon sperm DNA was incubated with the indicated units of TurboNuclease and another brand of nuclease at 37
o
C for 30 minutes in a buffer of 50 mM Tris-HCl, pH 8.0 and 1 mM MgCl
2
. DNA digestion was monitored by agarose gel.




TurboNuclease is purified through a proprietary process that achieves purity of >99%. TurboNuclease has no detectable protease activity.





TurboNuclease can be used to

reduce viscosity of cell lysate and reduce back pressure of column lo
ADI
ng

remove nucleic acid contamination from sample preparations, reduce nucleic acid contamination of Ni column purification

reduce smearing in SDS-PAGE when used with 10%SDS or Gel Lo
ADI
ng Dye to make whole cell lysate

reduce or prevent clumping of concentrated cells and thawed cells

replaces crude DNase I in many applications


To reduce viscosity of cell lysate, 10-500 units of TurboNuclease can be used for each gram of cell paste. Generally, adding TurboNuclease to cell lysate at 25 U/ml is sufficient to reduce lysate viscosity.

The efficiency of viscosity reduction may vary with buffers, cell types, and cell lysis methods used. Due to its high specific activity, the total amount TurboNuclease added is less than 0.1 ug/ml of lysate and will not complicate any down stream process.






Lare Scale Cell Lysis


Make fresh cold Lysis Buffer
Lysis Buffer should be a buffer in which the target protein is soluble.
The Lysis Buffer should be compat
IBL
e with downstream purification processes, e.g. minimal amount of EDTA or DTT if Ni column will be used.


Here is an example of Lysis Buffer
25 mM Tris-HCl, pH 8.0
500 mM NaCl
14 mM beta-mercaptoethanol
Detergent can be included for less soluble proteins or when protein solubility is unknown. 1% Triton X-100 has no effect on TurboNuclease activity.

TurboNuclease has the same activity in 150 mM NaCl or 500 mM NaCl and 400 mM imidazole.


Res
USP
end thawed cell paste in Lysis Buffer
Use 2-10 ml Lysis Buffer for each gram of cell paste. TurboNuclease can reduce the amount of Lysis Buffer used.
We routinely use 2 ml of lysis buffer for each gram of cell pellets.

Add TurboNuclease to 25 unit/ml
Protease inhibitors can be added at the same time.
If the lysis buffer contains EDTA or EGTA, add 10-fold more TurboNuclease.

Lyze cells by mechanical or chemical methods on ice or at room temperature
TurboNuclease also reduces the viscosity of lysate lyzed by microfluidizer.

Clear lysate by centrifugation for column lo
ADI
ng
The reduced viscosity makes it poss
IBL
e to clear the lysate at lower speed. 35,000g (~16,000 rpm) for 1 hour is sufficient.
Lysate can be loaded to "Crude" columns without clearance.


Parallel Lysis of Multiple Insect Cell Samples


Freeze cells pellets of 5-10 ml culture on dry ice briefly
Freeze and thaw facilitates lysis.

Thaw the frozen pellets and completely res
USP
end in ~1 ml Lysis Buffer with TurboNuclease

Transfer the cell s
USP
ension to a microtube and sit the tubes on a floater rack

Lyze cells using an Ultrasonic Cleaner with ice waterbath for 10 min
Ultrasonic Cleaner (many chemists use it) is much cheaper than probe sonicator. It costs a few hundreds US dollars for a new model and less than $100 for a used one or a jewelry cleaner from a consumer goods store. Ultrasonic Cleaner is much better and cheaper than those fancy multi-probe sonicators with the following advantages.


There is no cross-contamination since each sample is enclosed in a microtube.

The samples are always cold as long as ice is added in the water-bath.

There is no limit on the number of samples processed in parallel. A small Ultrasonic Cleaner can easily hold 48 samples

The lysate can be used for analyses of protein expression of whole cell lysate, soluble lysate, or affinity pull-down.

Eton Bio Lysis Buffer
25 mM Tris-HCl, pH 8.0
500 mM NaCl
20 mM Imidazole, pH 8.0
14 mM beta-mercaptoethanol
0.5% Triton X-100
25 units/ml TurboNuclease




Material Safety Data Sheet

Date:Oct 26,2015

1.

PRODUCT AND COMPANY IDENTIFICATION

1.1

Product identifier

Product name
: TurboNuclease

Catalog No
:1400010050,1400010100,1400010250

1.2

Relevant identified uses of the substance or mixture and uses advised against

For research use only.

1.3

Details of the supplier of the safety data sheet

Company:
Eton Bioscience
Inc

TollFree:1-800-758-1630

Tel:1-800-758-1630

Fax:1-800-507-2912

1.4

Emergency telephone number

1-800-758-1630


2.

HAZARDS IDENTIFICATION

2.1

Classification of the substance or mixture

Not a hazardous substance or mixture

2.2

GHS Label elements, including precautionary statements

Not a hazardous substance or mixture

2.3

Other hazards

None


3.

COMPOSITION/INFORMATION ON INGREDIENTS

3.1

Substances

Synonyms : Recombinant
Serratiamarcescens
extracellular endonuclease




CAS-No



EC-No.



Index-No



Classification



Concentration







Water





7732-18-5



231-791-2



-



-



>= 45 %







Glycerol

















56-81-5



200-289-5



-



-



50 %







Tris (hydroxymethyl) aminomethane free base





77-86-1



201-064-4



-



Xi, R36/37/38



0.6 %







Enzyme





-



-



-



-



<= 0.025 %







Magnesium chloride hexahydrate





7791-18-6



232-094-6



-



-



0.1 %







Sodium chloride





7647-14-5



231-598-3



-



-



0.3 %





For the full text of the R-phrases mentioned in this Section, see Section 16.


4.

FIRST AID MEASURES

4.1

Description of first aid measures

Eye contact

Check for and remove contact lenses and flush with copious amounts of water; assure adequate flushing by separating the eyelids with fingers; call a physician

Skin Contact

Flush with copious amounts of water; remove contaminated clothing and shoes; call a physician


Inhalation

Remove to fresh air
.
If not breathing give artificial respiration

Ingestion

If swallowed, never give anything by mouth to an unconscious person. Rinse mouth with water.

4.2

Most important symptoms and effects, both acute and delayed

No information available

4.3

Indication of any immediate medical attention and special treatment needed.

No information available


5.

FIRE FIGHTING MEASURES

5.1

Extinguishing media

Suitable extinguishing media

Water spray, alcohol-resistant foam, dry chemical or carbon dioxide

5.2

Special hazards arising from the substance or mixture

No information available

5.3

Advice for firefighters

Wear self-contained breathing apparatus and protective clothing to prevent contact with skin and eyes.

6.

ACCIDENTAL RELEASE MEASURES

6.1

Personal precautions,protective equipment and emergency procedures

Use full personal protective equipment. Evacuate personnel to safe areas. Avoid breathing vapors,mist or gas.

6.2

Environmental precautions

Prevent further leakage or spillage. Prevent product from entering drain.

6.3

Methods and material for containment and cleaning up

Keep in suitable, closed containers for disposal


7.

HANDLING AND STORAGE

7.1

Precautions for safe handling

Avoid skin/eye contact. Use protective equipment as needed. Wash contaminated clothing before reuse.

7.2

Conditions for safe storage,including any incompatibilities

Keep container tightly closed. Store at -80?C.

7.3

Specific end use(s)

No data available.


8.

EXPOSURE CONTROLS/PERSONAL INFORMATION

8.1

Control Parameters

This product contains no hazardous materials with occupational exposure limits.

8.2

Exposure controls

Engineering controls

Provide shower and eye wash station

Personal protective equipment




Eye Protection



Wear safety goggles





Skin Protection



Wear protective clothing





Hand Protection



Wear protective gloves





Respiratory protection



Wear NIOSH/MSHA approved respirators





9.

PHYSICAL AND CHEMICAL PROPERTIES

9.1

Information on basic physical and chemical properties




Appearance



Clear, colorless liquid





pH



8.0





Melting point



No data available





Flash point



No data available





Boiling point



No data available





Ignition temperature



No data available





Lower explosion limit



No data available





Upper explosion limit



No data available





Water solubility



Soluble





9.2

Other safety information

No data available.


10.

ST
ABI
LITY AND REACTIVITY

10.1
Reactivity

No data available

10.2
Chemical st
ABI
lity

Stable under recommended storage conditions.

10.3
Possibility of hazardous reactions

No data available.

10.4
Conditions to avoid

No data available

10.5
Incompat
IBL
e materials

Strong oxidizing agents

10.6
Hazardous decomposition products

Hazardous decomposition products formed under fire conditions: carbon oxides


11.

TOXICOLOGICAL INFORMATION

11.1
Information on toxicological effects

Acute toxicity

No data available


Irritation and corrosion

No data available


Sensitisation

Prolonged or repeated exposure may cause allergic reactions in certain sensitive individuals


Chronic exposure

IARC: No component of this product present at levels greater than or equal to 0.1% is identified as probable, poss
IBL
e or confirmed human carcinogen by IARC.


Signs and Symptoms o Exposure

Prolonged or repeated exposure can cause:Nausea,Headache,Vomiting


Potential Health Effects




Inhalation



May be harmful if inhaled. May cause respiratory tract irritation





Skin



May be harmful if absorbed through skin. May cause skin irritation





Eyes



May cause eye irritation





Ingestion



May be harmful if swallowed.





Target organs



Kidney





12.

ECOLOGICAL INFORMATION

12.1
Toxicity

Avoid release into environment

12.2
Persistence and degrad
ABI
lity

No data available

12.3
.
Bioaccumulative potential

No data available

12.4
Mobility in soil

No data available



12.5
Results of PBT and vPvB assessment

No data available

12.6
Other adverse effects








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