Ultra Nuclease GMP-urwego
Injangwe No: HC2016A
UltraNuclease GMP-igaragazwa kandi igasukurwa muri Escherichia coli (E.coli) ikozwe na genetique kandi yateguwe munsi ya GMP.Irashobora kugabanya ububobere bwa selile ndengakamere na lysate ya selile mubushakashatsi bwa siyanse, kongera imikorere ya protein no kongera ubushakashatsi bwa protein.Igicuruzwa kirashobora kandi kugabanya ibisigisigi bya acide nucleic kugeza kuri pg-urwego, kuzamura imikorere numutekano wibicuruzwa bikomoka kubinyabuzima birimo kweza virusi, gukora inkingo, hamwe n’imiti ya poroteyine / polysaccharide.Byongeye kandi, ibicuruzwa birashobora no gukoreshwa kugirango hirindwe ko ingirabuzimafatizo ziva mu maraso ya muntu (PBMC) mu kuvura ingirabuzimafatizo no guteza imbere inkingo.
UltraNuclease itangwa muburyo bwa reagent sterisile, yavuzwe muri buffer (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerine), igaragara nkamazi adafite ibara, abonerana.Ibicuruzwa byakozwe nibisabwa na GMP kandi bitangwa muburyo bwamazi.
Ibigize
UltraNuclease GMP-urwego (250 U / μL)
Ububiko
Ibicuruzwa byoherejwe hamwe na barafu yumye kandi birashobora kubikwa kuri -25 ℃ ~ -15 ° C mumyaka ibiri.
Niba ibicuruzwa byafunguwe kandi bibitswe kuri 4 ℃ mugihe kirenze icyumweru, turasaba kuyungururaibicuruzwa kugirango wirinde kwanduza mikorobe.
Ibisobanuro
| Ikirangantego | Recombinant E. coli hamwe na UltraNuclease gene |
| Uburemere bwa molekile | 26.5 kDa |
| ingingo ya soelectric | 6.85 |
| Isuku | ≥99% (SDS-PAGE) |
| Ububiko | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerine |
|
Igisobanuro | Igisobanuro cyigikorwa kimwe (U) nubunini bwa enzyme yakoreshejwehindura agaciro ko kwinjiza ΔA260 kuri 1.0 muminota 30 muri 2. 625 mLSisitemu ya reaction kuri 37 ℃ hamwe na pH ya 8.0 (bihwanye no gusya byuzuye37 μg intanga za salmon ADN muri oligonucleotide). |
Amabwiriza
1. Icyitegererezo Icyegeranyo
Utugingo ngengabuzima: kura hagati, koza selile hamwe na PBS, hanyuma ukureho ndengakamere.
Guhagarika ingirabuzimafatizo: gukusanya selile ukoresheje centrifugation, koza selile hamwe na PBS, centrifuge kuri 6.000rpm kuminota 10, kusanya pellet.
Escherichia coli: gukusanya bagiteri ukoresheje centrifugation, koza rimwe hamwe na PBS, centrifuge kuri 8,000rpm kuminota 5, hanyuma ukusanyirize pellet.
2. Uburyo bwo kuvura
Koresha pellet selile zegeranijwe hamwe na lysis buffer ku kigereranyo cya misa (g) nubunini (mL) 1: (10-20), cyangwa ukoresheje uburyo bwa mehaniki cyangwa chimique kurubura cyangwa mubushyuhe bwicyumba (1g ya pellet selile irimo hafi
Ingirabuzimafatizo 109).
3. Kuvura Enzyme
Ongeramo 1-5mM MgCl kuri sisitemu ya reaction hanyuma uhindure pH kuri 8-9.
Ongeramo UltraNuclease ukurikije igipimo cya Units 250 kugirango igogwe 1 g ya pellet selile, incubate kuri 37 ℃ muminota irenga 30.Nyamuneka reba ifishi "Yasabwe Kwitabira Igihe" kugirango uhitemoigihe cyo kwivuza.
4. Indengakamere Icyegeranyo
Centrifuge saa 12,000 rpm muminota 30 hanyuma ukusanyirize ndengakamere.
Icyitonderwa: Niba igisubizo ari acide cyangwa alkaline, cyangwa kirimo umunyu mwinshi, imyanda, cyangwadenaturants, nyamuneka wongere dosiye ya enzyme cyangwa wongere igihe cyo kuvura ukurikije.
Basabwe reaction conditions
| Parameter | Imiterere myiza | Umuyoboro mwiza |
| Mg² + Kwibanda | 1-5 mM | 1-10 mM |
| pH | 8-9 | 6-10 |
| Ubushyuhe | 37 ℃ | 0-42 ℃ |
| Kwibanda kwa DTT | 0-100 mM | > 0 mM |
| Kwibanda kwa Mercaptoethanol | 0-100 mM | > 0 mM |
| Kwibanda kuri Monovalent | 0-20 mM | 0-150 mM |
| Kwibanda kuri fosifate | 0-10 mM | 0-100 mM |
Basabwe Igisubizo Igihe (37 ℃, 2 mM Mg, +, pH 8.0)
| UltraNuclease Amafaranga (Kwibanda kwanyuma) | Igihe cyo Kwitwara |
| 0.25 U / mL | > 10 h |
| 2.5 U / mL | > 4h |
| 25 U / mL | 30 min |
Inyandiko:
Nyamuneka nyamuneka wambare PPE ikenewe, ikote rya laboratoire hamwe na gants, kugirango ubuzima bwawe n'umutekano wawe!
