DNAase (Synonyms: DNase I) |
Catalog No.GC19804
One-Click Copy Product Info
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≥2,000 Kunitz units/mg dry weight
DNAase (Deoxyribonuclease I, DNase I) (EC 3.1.21.1) is an endonuclease that can cleave single-stranded and double-stranded DNA, hydrolyzing phosphodiester bonds to form two products with 5'-phosphate termini and 3'-OH termini. DNase I is often used to enzymatically separate residual DNA in the digestion system of cells, or to study DNA-protein interactions using DNase I footprinting.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 9003-98-9
Sample solution is provided at 25 µL, 10mM.
DNAase (Deoxyribonuclease I, DNase I) (EC 3.1.21.1) is an endonuclease that can cleave single-stranded and double-stranded DNA, hydrolyzing phosphodiester bonds to form two products with 5'-phosphate termini and 3'-OH termini [1]. DNase I is often used to enzymatically separate residual DNA in the digestion system of cells, or to study DNA-protein interactions using DNase I footprinting [2, 3]. DNase I digestion of cells can increase cell viscosity without destroying cell integrity [4]. The activity of DNase I is strictly dependent on Ca2+ and is activated by Mg2+ or Mn2+ ions [5]. During the large-scale production of mRNA, the transcription template needs to be removed after transcription is completed. DNase I can randomly decompose single-stranded or double-stranded DNA to the same extent to generate oligonucleotides with 5'-P ends. Under Mg2+ conditions, DNase I can randomly shear double-stranded DNA[6, 7]. This DNAase product is derived from the bovine pancreas, the specific activity is ≥2000 Kunit Units/mg protein, stable at pH 7-8, and has a molecular weight of approximately 31 kDa.
Activity definition: DNase I catalyzes substrate DNA at 25°C and pH 5.0, resulting in a change of 0.001 in ∆A260 per milliliter per minute, which is defined as one Kunitz unit.
In vivo, DNase I (1000 U/500 μL) was injected intravenously into mice undergoing tenotomy, disrupting the DNA scaffold of neutrophil extracellular traps (NETs) and increasing neutrophil infiltration after injury[8].
References:
[1]Lauková L, Konečná B, Janovičová Ľ, et al. Deoxyribonucleases and their applications in biomedicine[J]. Biomolecules, 2020, 10(7): 1036.
[2]Miersch C, Stange K, Röntgen M. Effects of trypsinization and combined trypsin, collagenase, and DNase digestion on liberation and in vitro function of satellite cells isolated from juvenile porcine muscles[J]. In Vitro Cellular & Developmental Biology-Animal, 2018, 54: 406-412.
[3]Antunes A, Golfieri G, Ferlicca F, et al. HexR controls glucose-responsive genes and central carbon metabolism in Neisseria meningitidis[J]. Journal of Bacteriology, 2016, 198(4): 644-654.
[4]Sapio R T, Burns C J, Pestov D G. Effects of hydrogen peroxide stress on the nucleolar redox environment and pre-rRNA maturation[J]. Frontiers in Molecular Biosciences, 2021, 8: 678488.
[5]Fraser M J, Tynan S J, Papaioannou A, et al. Endo-exonuclease of human leukemic cells: evidence for a role in apoptosis[J]. Journal of Cell Science, 1996, 109(9): 2343-2360.
[6]Suck D. DNA recognition by DNase I[J]. Journal of Molecular Recognition, 1994, 7(2): 65-70.
[7]Sam M. Mechanistic studies on chemical and biological nucleases[M]. University of California, Los Angeles, 2005.
[8]Agarwal S, Loder S J, Cholok D, et al. Disruption of neutrophil extracellular traps (NETs) links mechanical strain to post-traumatic inflammation[J]. Frontiers in immunology, 2019, 10: 2148.
This plan only provides a guide, please modify it to meet your specific needs.
1. Solution preparation
(1) Stock solution: Dissolve DNase I in 0.15 M NaCl solution, the optimal pH is 7.0-8.0, and can be prepared into a 5-10 mg/mL stock solution.
Note:
(i) Activator: DNase I requires MgCl2 for activation to achieve maximum activity.
(ii) Inhibitor: EDTA, EGTA, SDS.
(iii) Specificity: Double-strand specific endonuclease that degrades DNA.
Do not vortex when dissolving the product. Keep the enzyme on ice when using it. Store it at -20℃ after use. In addition, even if the stock solution of this product is frozen at -20℃, it may lose about
2. Preparation of myoblasts by digesting muscle tissue with DNase I[1] (from the literature, for reference only)
(1) Preparation of muscle tissue: Wash and mince the dissected muscle tissue thoroughly with scissors.
(2) Preparation of mixed enzyme solution: Prepare a mixture of trypsin solution (0.25%, GB10152), collagenase I (0.2%, GC19589) and DNase I (0.01%).
(3) Primary enzymatic digestion: Place the muscle tissue in a water bath, add the enzyme solution, and digest with stirring at 37°C for 60 minutes. Stop stirring and collect the cell slurry supernatant of the first digestion after 30 minutes.
(4) Stop digestion: Stop the digestion process by adding growth medium (αMEM Eagle, 20% FBS, 100 U/ml penicillin/streptomycin, 2.5 μg/ml amphotericin B, 0.05 mg/ml gentamicin).
(5) Secondary enzymatic digestion: Add fresh enzyme solution to the remaining muscle fragments and digest for another 30 minutes.
(6) Stop digestion: Add growth medium again to completely stop the digestion process, and combine the cell suspensions obtained from the two digestions.
(7) Cell suspension treatment: Use a 25 ml pipette to vigorously blow and homogenize the combined cell suspension. After washing, filter through gauze and a fine nylon mesh (20 μm), and enrich myoblasts by density gradient centrifugation.
3. Steps for preparing primary glioma cells using DNase I digestion of neural tissue [2] (from the literature, for reference only)
(1) Neural tissue preparation: Mechanically cut the tissue into small pieces.
(2) Enzymatic hydrolysis: Use collagenase I (GC19589) and DNase I for enzymatic hydrolysis at 37°C for 30 minutes.
(3) Washing: Wash cells with PBS buffer.
(4) Cell culture: Use DMEM medium to culture cells.
Note: This protocol provides reference steps for tissue digestion using DNase I. Please adjust the specific dosage according to the actual situation.
References:
[1] Miersch C, Stange K, Röntgen M. Effects of trypsinization and of a combined trypsin, collagenase, and DNase digestion on liberation and in vitro function of satellite cells isolated from juvenile porcine muscles[J]. In Vitro Cellular & Developmental Biology-Animal, 2018, 54: 406-412.
[2] Sim J M, Park J M, Kim S, et al. Association of tim-3/gal-9 axis with NLRC4 inflammasome in glioma malignancy: Tim-3/Gal-9 induce the NLRC4 inflammasome[J]. International Journal of Molecular Sciences, 2022, 23(4): 2028.
| Cas No. | 9003-98-9 | SDF | |
| Synonyms | DNase I | ||
| Formula | M.Wt | ~ 31KDa | |
| Solubility | DMSO : 43.48 mg/mL (adjust pH to 3 with 1M HCl); H2O : < 100mg/mL | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
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Biological Activity: ≥2000 Kunitz units/mg protein Appearance: A solid
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 28 reference(s) in Google Scholar.)