Ultra Nuclease GMP-ibakala
Inombolo yekati: HC2016A
I-UltraNuclease GMP-grade ichazwa kwaye ihlanjululwe kwi-Escherichia coli (E.coli) nge-genetically engineered kwaye ilungiswe phantsi kweemeko ze-GMP.Inokunciphisa i-viscosity ye-cell supernatant kunye ne-cell lysate kuphando lwezenzululwazi, ukwandisa ukusebenza kweprotheyini yokucoca kunye nokuphucula uphando olusebenzayo lweprotheni.Imveliso inokunciphisa iintsalela ze-nucleic acid ezihlala kwi-pg-grade, ukuphucula ukusebenza kunye nokhuseleko lweemveliso zebhayoloji zezicelo ezibandakanya ukucocwa kwentsholongwane, ukuveliswa kwesitofu sokugonya, kunye neprotheyini / i-polysaccharide yokuvelisa amayeza.Ngaphandle koko, imveliso ingasetyenziselwa ukuthintela ukudityaniswa kweeseli zegazi ze-mononuclear zegazi (PBMC) kunyango lweeseli kunye nophuhliso lwesitofu.
I-UltraNuclease inikwe ngohlobo lwe-reagent entsholongwane, eluted kwi-buffer (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), kunye nenkangeleko yolwelo olungenambala, olucacileyo.Le mveliso iveliswa ngokweemfuno zenkqubo ye-GMP kwaye inikezelwe ngendlela engamanzi.
Amacandelo
I-UltraNuclease GMP-grade (250 U/μL)
Iimeko zokuGcina
Imveliso ithunyelwa ngeqhwa elomileyo kwaye ingagcinwa kwi -25 ℃ ~ -15 ° C iminyaka emibini.
Ukuba imveliso ivuliwe kwaye igcinwe kwi-4℃ ngaphezu kweveki, sincoma ukuhluzaimveliso yokuthintela ungcoliseko lwe-microbial.
Iinkcukacha
| Isivakalisi sokuvakalisa | I-Recombinant E. coli kunye ne-UltraNuclease gene |
| Ubunzima beMolekyuli | 26.5 kDa |
| indawo yombane | 6.85 |
| Ubunyulu | ≥99% (SDS-PAGE) |
| Isithinteli sokuGcina | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Inkcazelo yeyunithi | Inkcazo yeyunithi yoMsebenzi enye (U) sisixa se-enzyme esetyenziselwatshintsha ixabiso lokufunxa le-ΔA260 nge-1.0 kwimizuzu engama-30 kwi-2. 625 mLinkqubo yokusabela kwi-37℃ ene-pH ye-8.0 (elingana nokugaywa ngokupheleleyo37 μg i-salmon yesidoda se-DNA kwi-oligonucleotides). |
Imiyalelo
1. Isampuli Ingqokelela
Iiseli ezibambelelayo: susa okuphakathi, hlamba iiseli nge-PBS, kwaye ususe i-supernatant.
Iiseli ezimisiweyo: qokelela iiseli nge-centrifugation, hlamba iiseli nge-PBS, i-centrifuge kwi-6,000rpm i-10 min, qokelela i-pellet.
I-Escherichia coli: qokelela iibhaktheriya nge-centrifugation, hlamba kube kanye nge-PBS, i-centrifuge kwi-8,000rpm i-5 min, kwaye uqokelele i-pellet.
2. Unyango lweSampuli
Phatha iipellets zeseli eziqokelelweyo nge-lysis buffer kumlinganiselo wobunzima (g) ukuya kumthamo(mL)1:(10-20), okanye ngeendlela zoomatshini okanye zekhemikhali kumkhenkce okanye kubushushu begumbi (1g yepellet yeseli iqulathe malunga
Iiseli ezili-109).
3. Unyango lwe-Enzyme
Yongeza i-1-5mM MgCl kwinkqubo yokuphendula kwaye ulungelelanise i-pH kwi-8-9.
Yongeza i-UltraNuclease ngokomlinganiselo weeYunithi ezingama-250 ukwetyisa i-1 g yeepellets zeseli, zifukame ku-37℃ ngaphezu kwemizuzu engama-30.Nceda ujonge kwifomu ethi “Ixesha Lokuphendula eliCetyisiweyo” ukukhetha iixesha lonyango.
4. Onamandla Ingqokelela
I-Centrifuge kwi-12,000 rpm imizuzu engama-30 kwaye uqokelele i-supernatant.
Qaphela: Ukuba isisombululo sineasidi okanye i-alkaline, okanye iqulethe ityuwa eninzi, izinto zokucoca, okanyedenaturants, nceda wandise umthamo we-enzyme okanye wandise ixesha lonyango ngokufanelekileyo.
Conditi yokusabela ecetyiswayoons
| Ipharamitha | Imeko eNgcono | I-Conditior esebenzayo |
| Mg²+Ukugxila | 1-5 mm | 1-10 mm |
| pH | 8-9 | 6-10 |
| Ubushushu | 37℃ | 0-42℃ |
| Ugxininiso lweDTT | 0-100 mm | >0 mM |
| Ukugxilwa kweMercaptoethanol | 0-100 mm | >0 mM |
| Monovalent cation Concentration | 0-20 mm | 0-150 mm |
| I-Phosphate lon Concentration | 0-10 mm | 0-100 mm |
Kucetyiswa Ukusabela Ixesha (37℃, 2 mM Mg²+, pH 8.0)
| Isixa se-UltraNuclease (Uxinaniso lokugqibela) | Ixesha lokuphendula |
| 0.25 U/mL | >10 h |
| 2.5 U/mL | >4h |
| 25 U/mL | 30 imiz |
Amanqaku:
Nceda unxibe iPPE eyimfuneko, idyasi yelebhu kunye neeglavu, ukuqinisekisa impilo nokhuseleko lwakho!
