I-Ultra Nuclease GMP-grade
Inombolo yekati: HC2016A
I-UltraNuclease GMP-grade ichazwa futhi ihlanzwe ku-Escherichia coli (E.coli) ngokwakhiwa kofuzo futhi ilungiswe ngaphansi kwezindawo ze-GMP.Inganciphisa i-viscosity ye-cell supernatant kanye ne-cell lysate ocwaningweni lwesayensi, ikhulise ukusebenza kahle kokuhlanzwa kwamaprotheni futhi ithuthukise ucwaningo olusebenzayo lwamaprotheni.Umkhiqizo ungaphinda wehlise izinsalela ze-nucleic acid ezisingethe zibe pg-grade, uthuthukise ukusebenza nokuphepha kwemikhiqizo yebhayoloji yezinhlelo zokusebenza ezihlanganisa ukuhlanzwa kwegciwane, ukukhiqizwa kokugoma, kanye nokukhiqizwa kwemithi ye-protein/polysaccharide.Ngaphandle kwalokho, umkhiqizo ungase futhi usetshenziswe ukuvimbela ukunqwabelana kwamaseli egazi lomuntu we-peripheral mononuclear cell (PBMC) ekwelapheni kwamaseli nokuthuthukiswa komgomo.
I-UltraNuclease inikezwa ngesimo se-reagent ehlanzekile, ekhishwe ku-buffer (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), ebukeka njengoketshezi olungenambala, olusobala.Lo mkhiqizo ukhiqizwa yizidingo zenqubo ye-GMP futhi unikezwe ngendlela ewuketshezi.
Izingxenye
I-UltraNuclease GMP-grade (250 U/μL)
Izimo Zokugcina
Umkhiqizo uthunyelwa ngeqhwa elomile futhi ungagcinwa ku -25℃~-15°C iminyaka emibili.
Uma umkhiqizo uvuliwe futhi ugcinwe ku-4℃ isikhathi esingaphezu kweviki, sincoma ukuthi uhlungweumkhiqizo ukuvimbela ukungcoliswa kwamagciwane.
Imininingwane
| Expression Host | I-Recombinant E. coli enofuzo lwe-UltraNuclease |
| Isisindo samangqamuzana | 26.5 kDa |
| indawo kagesi | 6.85 |
| Ubumsulwa | ≥99% (SDS-PAGE) |
| Ibhafa Yesitoreji | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Incazelo Yeyunithi | Incazelo yeyunithi yoMsebenzi eyodwa (U) inani le-enzyme esetshenziselwashintsha inani lokumunca le-ΔA260 ngo-1.0 emaminithini angu-30 ku-2. 625 mLisistimu yokusabela ku-37℃ ene-pH engu-8.0 (okulingana nokugaya okuphelele37 μg i-salmon yesidoda se-DNA ibe yi-oligonucleotides). |
Iziyalezo
1. Isampula Iqoqo
Amaseli anamathelayo: khipha okuphakathi, geza amaseli nge-PBS, futhi ukhiphe amandla angaphezu kwavamile.
Amaseli amisiwe: qoqa amaseli nge-centrifugation, geza amaseli nge-PBS, i-centrifuge ku-6,000rpm imizuzu eyi-10, qoqa i-pellet.
I-Escherichia coli: qoqa amagciwane nge-centrifugation, geza kanye nge-PBS, i-centrifuge ku-8,000rpm imizuzu emi-5, bese uqoqa i-pellet.
2. Ukwelashwa okuyisampula
Phatha ama-cell pellets aqoqiwe nge-lysis buffer ngesilinganiso se-mass(g) nevolumu(mL)1:(10-20), noma ngezindlela zemishini noma zamakhemikhali eqhweni noma ekushiseni kwegumbi (1g ye-cell pellet iqukethe cishe
109 amaseli).
3. Ukwelashwa Kwe-enzyme
Engeza i-1-5mM MgCl kusistimu yokusabela futhi ulungise i-pH ibe ngu-8-9.
Engeza i-UltraNuclease ngokwesilinganiso sama-Units angu-250 ukuze ugaye i-1 g yama-cell pellets, ufukamele ku-37℃ ngaphezu kwemizuzu engama-30.Sicela ubheke ifomu elithi “Isikhathi Sokusabela Esinconyiwe” ukuze ukhetheubude besikhathi sokwelashwa.
4. Onamandla Iqoqo
I-Centrifuge ku-12,000 rpm imizuzu engu-30 futhi iqoqe i-supernatant.
Qaphela: Uma isixazululo sine-acidic noma sine-alkaline, noma siqukethe usawoti omningi, okokuhlanza, nomaama-denaturants, sicela wandise umthamo we-enzyme noma wandise isikhathi sokwelashwa ngokufanele.
I-conditi yokusabela enconyiweons
| Ipharamitha | Isimo Esilungile | I-Conditior Esebenzayo |
| I-Mg²+Concentration | 1-5 mM | 1-10 mm |
| pH | 8-9 | 6-10 |
| Izinga lokushisa | 37℃ | 0-42℃ |
| I-DTT Concentration | 0-100 mM | >0 mM |
| I-Mercaptoethanol Concentration | 0-100 mM | >0 mM |
| I-Monovalent Cation Concentration | 0-20 mm | 0-150 mM |
| I-Phosphate lon Concentration | 0-10 mm | 0-100 mM |
Kunconyiwe Ukusabela Isikhathi (37℃, 2 mM Mg²+, pH 8.0)
| Inani le-UltraNuclease (Ukugxila Kokugcina) | Isikhathi sokuphendula |
| 0.25 U/mL | >10h |
| 2.5 U/mL | > 4h |
| 25 U/mL | 30 imiz |
Amanothi:
Sicela ugqoke i-PPE edingekayo, ijazi lelebhu kanye namagilavu, ukuze uqinisekise impilo nokuphepha kwakho!
