Rnase A
Ribonuclease A(RNaseA) i polypeptide ine tambo imwe ine 4 disulfide bond ine huremu hwema molecular hunosvika 13.7 kDa.RNase Ais an endoribonuclease iyo inonyanya kushatisa imwechete-stranded RNA paC uye U zvakasara.Kunyanya, iyo cleavage inoona phosphodiester bond inoumbwa ne5'-ribose ye nucleotide uye phosphate boka pa 3'-ribose yepedyo pyrimidine nucleotide, kuitira kuti 2,3'-Cyclic phosphates igadzirwe kune inoenderana ne3. 'nucleoside phosphates (semuenzaniso,pG-pG-pC-pA-pG inochekwa neRNase A kugadzira pG-pG-pCp uye A-PG).RNase A ndiyo inonyanya kushanda mukutsemura ine tambo imwe chete RNA.Yakakurudzirwa kushanda kusungirirwa ndeye 1- 100 μ G/mL, inoenderana neakasiyana maitiro masisitimu.Munyu wakaderera (0-100 mM NaCl) unogona kushandiswa kucheka RNA ine tambo imwe chete, ine mbiri-yakapetwa RNA, uye RNA cheni dzakaumbwa neRNA-DNA hybridization.
Nekudaro, pakuwanda kwemunyu (≥0.3 M), RNAse A inongotsemura imwe-stranded RNA.
RNase A inonyanya kushandiswa kubvisa RNA panguva yekugadzirira plasmid DNA kana genomic DNA.Kunyangwe kana kwete DNase iri kushanda panguva yekugadzirira inogona kukanganisa maitiro.Nzira yechinyakare yekubikira mumvura yekugezera inogona kushandiswa kumisa basa reDNase.Ichi chigadzirwa hachina DNase uye protease, uye haidi kurapwa kwekupisa isati yashandiswa.Uye zvakare, chigadzirwa ichi chinogona zvakare kushandiswa mumamorekuru biology kuyedza senge RNase kudzivirira kuongororwa uye RNA kutevedzana kuongororwa.
Kuchengetedza mamiriro
Chigadzirwa chacho chinofanira kuchengetwa pa -25 ℃ ~ - 15 ℃ kwemakore maviri.
Zvinotsanangurwa
| Chitarisiko | Upfu |
| Quantity | 100mg / 1g |
| Chigadzirwa mhando | RNase A |
Mirayiridzo
Iyi ndiyo imwe yedzakajairika nzira dzekugadzirira RNase A kuchengetedza mhinduro.Inogonawo kugadzirirwanedzimwe nzira maererano nemaitiro echinyakare murabhoritari kana mabhuku ereferenzi (sengekunyungudika zvakananga mu10 mM Tris-HCl, pH 7.5 kana Tris-NaCl mhinduro)
1. Shandisa 10 mM sodium acetate (pH 5.2) kugadzirira 10 mg / mL yeRNase A yekuchengetedza mhinduro.
2. Kupisa pa100 ℃ kwemaminitsi gumi nemashanu.
3. Kutonhoresa kune tembiricha yekamuri, wedzera 1/10 vhoriyamu ye1 M Tris-HCl (pH 7.4), gadzirisa pH yayo kusvika 7.4 (yesomuenzaniso, wedzera 500 mL ye10 mg/mL RNase storage solution 1 M Tris-HCl, pH7.4).
4. Yakavharwa pa -20 ℃ kuitira kuchengetedza nechando, iyo inogona kugadzikana kusvika kumakore maviri.
[Zvinyorwa]:
Paunenge uchibika mhinduro yeRNaseA pasi pemamiriro asina kwaakarerekera, kunaya kweRNase kunoumba;Ifashaidze pane yakaderera pH, uye kana paine kunaya, inogona kucherechedzwa, iyo inogona kukonzerwa nekuvapo kweprotein tsvina.Kana sediment yawanikwa mushure mekuvira, tsvina inogona kubviswa ne-high-speed centrifugation (13000rpm), uyezve pasi-packed kuti ichengetedzwe nechando.
Notes
Ndokumbira upfeke PPE inodiwa, jasi rakadaro uye magurovhosi, kuve nechokwadi chehutano hwako uye kuchengetedzeka.
