Rnase A
Ribonuclease A (RNaseA) i polypeptide ine tambo imwe chete ine 4 disulfide bond ine molecular huremu hunosvika 13.7 kDa.RNase A inonzi endoribonuclease iyo inonyanya kushatisa imwe-stranded RNA paC uye U zvakasara.Kunyanya, iyo cleavage inoziva phosphodiester bond inoumbwa ne5'-ribose ye nucleotide uye phosphate boka pane 3'-ribose yepedyo pyrimidine nucleotide, kuitira kuti iyo 2, 3'-Cyclic phosphates inogadziriswa hydrolyzed kune inoenderana 3. nucleoside phosphates (semuenzaniso pG-pG-pC-pA-pG inotsemurwa neRNase A kugadzira pG-pG-pCp uye A-PG).RNase A ndiyo inonyanya kushanda mukutsemura ine tambo imwe chete RNA.Yakakurudzirwa kushanda kusungirirwa ndeye 1-100μG/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 chete-stranded RNA.
RNase A inonyanya kushandiswa kubvisa RNA panguva yekugadzirira plasmid DNA kana genomic DNA.Kuti kana kwete DNase inoshanda panguva yekugadzirira inogona kuchinja nyore nyore maitiro.Nzira yechinyakare yekubikira mumvura yekugezera inogona kushandiswa kumisa basa reDNase.Ichi chigadzirwa hachina DNase uye protease, uye haidi kurapwa kwekupisa isati yashandiswa.Pamusoro pezvo, chigadzirwa ichi chinogona zvakare kushandiswa mumamolecular biology kuyedza senge RNase kudzivirira kuongororwa uye RNA kutevedzana kuongororwa.
Storage Conditions
Chigadzirwa chacho chinogona kuchengetwa pa -25～-15 ℃, inoshanda kwemakore maviri.
Mirayiridzo
Iyi ndiyo imwe yedzakajairika nzira dzekugadzirira RNase A kuchengetedza mhinduro.Inogonawo kugadzirwa nedzimwe 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 (yesemuenzaniso, wedzera 500 ml ye10g/ml RNase yekuchengetedza mhinduro 1M Tris-HCL, PH7.4)
4. Sub-package at -20℃ yekuchengetedza nechando, inogona kugadzikana kusvika kumakore maviri.
[Zvinyorwa]: Kana uchibika mhinduro yeRNaseA pasi pemamiriro asina kwaakarerekera, kunaya kweRNase kuchaumba;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-yakarongedzerwa kuchengetedza kutonhora.
Ruzivo rwechigadzirwa
| Mashoko anoreva zvakafanana | Ribonuclease I;Pancreatic ribonuclease;Ribonuclease 3'-pyrimidnooligonucleotidohydrolase;Rnase A;Endoribonulcease I |
| CAS Nha. | 9001-99-4 |
| Chitarisiko | White lyophilized powder |
| Molecular uremu | ~ 13.7kDa (amino acid kutevedzana) |
| Ph Kukosha | 7.6 (chiitiko chiyero 6-10) |
| Yakakodzera Tembiricha | 60 ℃ (Basa renji 15-70 ℃) |
| Activating agent | Na2+.K+ |
| Inhibitor | Rnase Inhibitor |
| Inactivation nzira | Haikwanise kuvharwa nekudziya, zvinokurudzirwa kushandisa centrifuge column |
| Origin | Bovine |
| Solubility | Soluble mumvura (10mg/ml) |
| Kurasikirwa pakaoma | ≤5.0% |
| Enzyme chiitiko | ≥60 Kunitz zvikamu/mg |
| Isoelectric point | 9.6 |
Notes
Nekuchengetedza uye hutano hwako, ndapota pfeka majasi erabhu uye magurovhosi anoraswa kuti ushande.
