Ultra Nuclease GMP-grade
Nomoro ea katse: HC2016A
UltraNuclease GMP-grade e hlahisoa le ho hloekisoa ho Escherichia coli (E.coli) ka liphatsa tsa lefutso le ho lokisoa tlas'a maemo a GMP.E ka fokotsa viscosity ea cell supernatant le cell lysate lipatlisisong tsa mahlale, ea eketsa ts'ebetso ea tlhoekiso ea protheine le ho ntlafatsa lipatlisiso tsa ts'ebetso ea protheine.Sehlahisoa se ka boela sa fokotsa masala a acid a nucleic acid ho pg-grade, ho ntlafatsa ts'ebetso le polokeho ea lihlahisoa tsa likokoana-hloko tse sebelisoang ho kenyeletsa ho hloekisa kokoana-hloko, tlhahiso ea ente, le tlhahiso ea meriana ea liprotheine / polysaccharide.Ntle le moo, sehlahisoa se ka sebelisoa hape ho thibela ho bokellana ha lisele tsa mali tsa "pharipheral blood mononuclear" (PBMC) kalafong ea lisele le nts'etsopele ea ente.
UltraNuclease e fanoe ka mokhoa oa reagent e hloekisitsoeng, e hlahisitsoeng ka har'a buffer (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), e nang le ponahalo ea metsi a se nang 'mala, a bonaletsang.Sehlahisoa sena se hlahisoa ke litlhoko tsa ts'ebetso ea GMP mme se fanoa ka mokhoa oa mokelikeli.
Likaroloana
UltraNuclease GMP-grade (250 U/μL)
Maemo a polokelo
Sehlahisoa se romelloa ka leqhoa le omileng 'me se ka bolokoa ho -25 ℃~-15 ° C ka lilemo tse peli.
Haeba sehlahisoa se butsoe 'me se bolokiloe ho 4 ℃ nako e fetang beke, re khothaletsa ho sefasehlahisoa ho thibela tšoaetso ea likokoana-hloko.
Litlhaloso
| Expression Host | Recombinant E. coli e nang le lefutso la UltraNuclease |
| Boima ba Molek'hule | 26.5 kDa |
| ntlha ea motlakase | 6.85 |
| Bohloeki | ≥99% (SDS-PAGE) |
| Buffer ea polokelo | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Tlhaloso ea Yuniti | Tlhaloso ea karolo e le 'ngoe ea Ketsahalo (U) ke palo ea enzyme e sebelisoang hofetola boleng ba ho monya ba ΔA260 ka 1.0 ka metsotso e 30 ka 2. 625 mLsistimi ea karabelo ho 37 ℃ ka pH ea 8.0 (e lekanang le tšilo e felletseng ea37 μg ea peō ea salmon ea DNA ho oligonucleotides). |
Litaelo
1. Mohlala Pokello
Lisele tse khomarelang: tlosa sehare, hlatsoa lisele ka PBS, 'me u tlose matla a phahametseng tlhaho.
Lisele tse emisitsoeng: bokella lisele ka centrifugation, hlatsoa lisele ka PBS, centrifuge ho 6,000rpm bakeng sa 10 min, bokella pellet.
Escherichia coli: bokella libaktheria ka centrifugation, hlatsoa hang ka PBS, centrifuge ho 8,000rpm bakeng sa 5 min, 'me u bokelle pellet.
2. Mohlala oa Kalafo
Tšoara li-pellets tsa lisele tse bokelitsoeng ka lysis buffer ka karo-karolelano ea boima(g) ho volume(mL)1:(10-20), kapa ka mekhoa ea mochini kapa ea lik'hemik'hale holim'a leqhoa kapa mocheso oa kamore (1g ea pellet ea sele e na le hoo e ka bang
109 lisele).
3. Kalafo ea Enzyme
Kenya 1-5mM MgCl ho sistimi ea karabelo 'me u fetole pH ho 8-9.
Eketsa UltraNuclease ho ya ka karo-karolelano ya Diyuniti tse 250 ho sila 1 g ya di-pellets tsa disele, e kenye ka 37℃ ho feta metsotso e 30.Ka kopo sheba foromo ea "Recommended Reaction Time" ho khethanako ya kalafo.
4. Motho ea phahameng ka ho fetisisa Pokello
Centrifuge ka 12,000 rpm bakeng sa metsotso e 30 'me u bokelle supernatant.
Tlhokomeliso: Haeba tharollo e le acidic kapa alkaline, kapa e na le letsoai le lengata, litlolo, kapali-denaturants, ka kopo eketsa litekanyetso tsa enzyme kapa eketsa nako ea kalafo ka nepo.
Recommended reaction conditions
| Paramethara | Boemo bo nepahetseng | E Atlehang Conditior |
| Mg²+Concentration | 1-5 mmM | 1-10 mmM |
| pH | 8-9 | 6-10 |
| Mocheso | 37℃ | 0-42 ℃ |
| Khokahano ea DTT | 0-100 mmM | >0 mM |
| Mahlohonolo a Mercaptoethanol | 0-100 mmM | >0 mM |
| Monovalent Cation Concentration | 0-20 mmM | 0-150 mmM |
| Phosphate lon Concentration | 0-10 mmM | 0-100 mmM |
E khothalelitsoe Boitšoaro Nako (37℃, 2 mM Mg²+, pH 8.0)
| Chelete ea Nuclease (Tlhahiso ea ho Qetela) | Nako ea ho itšoara |
| 0.25 U/mL | >10 h |
| 2.5 U/mL | >4h |
| 25 U/mL | 30 mets |
Lintlha:
Ka kopo, roala PPE e hlokahalang, jase e joalo ea lab le liatlana, ho netefatsa bophelo bo botle le polokeho ea hau!
