Everest Biotech/TurboNuclease 50,000 Units/1400010050/50,000 Units
Protein Type:EndonucleaseSource:E. coliUnit Definition:One unit of TurboNuclease converts 1.0 OD260 of salmon sperm DNA into acid-soluble nucleotides in 30 minutes at 37oC in a reaction buffer of 50 mM Tris-HCl, pH 8.0 and 1 mM MgCl2. This corresponds to complete digestion of 50 µg of salmon sperm DNA into oligonucleotides. TurboNuclease has similar specific activity as BenzonaseStorage:Store TurboNuclease at -20oC.Endotoxin Level:< 0.1 EU / 1,000 units of TurboNuclease as determined by the Endo-Safe LAL Assay. Formulation:
250 units/µl in 50 mM Tris-HCl, pH 8.0, 50 mM NaCl, 5 mM MgCl2 and 50% Glycerol
TurboNuclease Activity and Purity:
| 50 µg of salmon sperm DNA was incubated with the indicated units of TurboNuclease and another brand of nuclease at 37oC for 30 minutes in a buffer of 50 mM Tris-HCl, pH 8.0 and 1 mM MgCl2. 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 loading 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 Loading 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 compatible with downstream purification processes, e.g. minimal amount of EDTA or DTT if Ni column will be used.Here is an example of Lysis Buffer25 mM Tris-HCl, pH 8.0500 mM NaCl14 mM beta-mercaptoethanolDetergent 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. Resuspend 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 loading The reduced viscosity makes it possible 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 resuspend in ~1 ml Lysis Buffer with TurboNuclease Transfer the cell suspension 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 Buffer25 mM Tris-HCl, pH 8.0500 mM NaCl20 mM Imidazole, pH 8.014 mM beta-mercaptoethanol0.5% Triton X-10025 units/ml TurboNuclease Material Safety Data SheetDate:Oct 26,20151. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name: TurboNucleaseCatalog No:1400010050,1400010100,14000102501.2 Relevant identified uses of the substance or mixture and uses advised againstFor research use only.1.3 Details of the supplier of the safety data sheetCompany:Eton Bioscience IncTollFree:1-800-758-1630Tel:1-800-758-1630Fax:1-800-507-29121.4 Emergency telephone number1-800-758-16302. HAZARDS IDENTIFICATION2.1 Classification of the substance or mixtureNot a hazardous substance or mixture2.2 GHS Label elements, including precautionary statementsNot a hazardous substance or mixture2.3 Other hazardsNone3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms : Recombinant Serratiamarcescensextracellular 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 MEASURES4.1 Description of first aid measuresEye contactCheck for and remove contact lenses and flush with copious amounts of water; assure adequate flushing by separating the eyelids with fingers; call a physicianSkin ContactFlush with copious amounts of water; remove contaminated clothing and shoes; call a physicianInhalationRemove to fresh air. If not breathing give artificial respirationIngestionIf swallowed, never give anything by mouth to an unconscious person. Rinse mouth with water.4.2 Most important symptoms and effects, both acute and delayedNo information available4.3 Indication of any immediate medical attention and special treatment needed.No information available5. FIRE FIGHTING MEASURES5.1 Extinguishing mediaSuitable extinguishing mediaWater spray, alcohol-resistant foam, dry chemical or carbon dioxide5.2 Special hazards arising from the substance or mixtureNo information available5.3 Advice for firefightersWear self-contained breathing apparatus and protective clothing to prevent contact with skin and eyes.6. ACCIDENTAL RELEASE MEASURES6.1 Personal precautions,protective equipment and emergency proceduresUse full personal protective equipment. Evacuate personnel to safe areas. Avoid breathing vapors,mist or gas.6.2 Environmental precautionsPrevent further leakage or spillage. Prevent product from entering drain.6.3 Methods and material for containment and cleaning upKeep in suitable, closed containers for disposal7. HANDLING AND STORAGE7.1 Precautions for safe handlingAvoid skin/eye contact. Use protective equipment as needed. Wash contaminated clothing before reuse.7.2 Conditions for safe storage,including any incompatibilitiesKeep container tightly closed. Store at -80?C.7.3 Specific end use(s)No data available.8. EXPOSURE CONTROLS/PERSONAL INFORMATION8.1 Control ParametersThis product contains no hazardous materials with occupational exposure limits.8.2 Exposure controlsEngineering controlsProvide shower and eye wash stationPersonal 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 PROPERTIES9.1 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 informationNo data available.10. STABILITY AND REACTIVITY10.1ReactivityNo data available10.2Chemical stabilityStable under recommended storage conditions.10.3Possibility of hazardous reactionsNo data available.10.4Conditions to avoidNo data available10.5Incompatible materialsStrong oxidizing agents10.6Hazardous decomposition productsHazardous decomposition products formed under fire conditions: carbon oxides11. TOXICOLOGICAL INFORMATION11.1Information on toxicological effectsAcute toxicityNo data availableIrritation and corrosionNo data availableSensitisationProlonged or repeated exposure may cause allergic reactions in certain sensitive individualsChronic exposureIARC: No component of this product present at levels greater than or equal to 0.1% is identified as probable, possible or confirmed human carcinogen by IARC.Signs and Symptoms o ExposureProlonged or repeated exposure can cause:Nausea,Headache,VomitingPotential 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 INFORMATION12.1ToxicityAvoid release into environment12.2Persistence and degradabilityNo data available12.3.Bioaccumulative potentialNo data available12.4Mobility in soilNo data available12.5Results of PBT and vPvB assessmentNo data available12.6Other adverse effects
