Rnase A
I-Ribonuclease A (i-RNaseA) yi-polypeptide ene-single-stranded equkethe i-4 disulfide bond kunye ne-molecular weight malunga ne-13.7 kDa.I-RNase A yi-endoribonuclease eyehlisa ngokukodwa i-RNA enemisonto enye kwi-C kunye neentsalela ze-U.Ngokukodwa, i-cleavage iyaqaphela ibhondi ye-phosphodiester eyenziwe yi-5'-ribose ye-nucleotide kunye neqela le-phosphate kwi-3'-ribose ye-nucleotide esondeleyo ye-pyrimidine, ukwenzela ukuba i-2, i-3'-Cyclic phosphates ifakwe kwi-hydrolyzed kwi-3 ehambelanayo. Inucleoside phosphates (umz. pG-pG-pC-pA-pG icandwa yiRNase A ukuvelisa i-pG-pG-pCp kunye ne-A-PG).I-RNase A yeyona nto isebenzayo ekucakeni i-RNA enomsonto omnye.Ugxininiso lokusebenza olucetyiswayo yi-1-100μG/mL, iyahambelana neenkqubo ezahlukeneyo zokusabela.I-concentration yetyuwa ephantsi (i-0-100 mM NaCl) ingasetyenziselwa ukusika i-RNA enye, i-RNA ene-double-stranded, kunye ne-RNA ye-RNA eyenziwe yi-RNA-DNA hybridization.Nangona kunjalo, kugxininiso lwetyuwa ephezulu (≥0.3 M), i-RNase A iqhawula kuphela i-RNA enomsonto omnye.
I-RNase A isetyenziswa ngokuqhelekileyo ukususa i-RNA ngexesha lokulungiswa kwe-plasmid DNA okanye i-DNA ye-genomic. Ingaba okanye i-DNase ingasebenzi ngexesha lenkqubo yokulungiselela inokuchaphazela ngokulula ukusabela.Indlela yendabuko yokubilisa kwindawo yokuhlambela amanzi ingasetyenziselwa ukungasebenzi komsebenzi we-DNase.Le mveliso ayinayo i-DNase kunye ne-protease, kwaye ayifuni unyango olushushu ngaphambi kokusetyenziswa.Ukongeza, le mveliso inokusetyenziswa kuvavanyo lwebhayoloji yemolekyuli njengohlalutyo lokhuseleko lwe-RNase kunye nohlalutyo lokulandelelana kwe-RNA.
Iimeko zokuGcina
Imveliso inokugcinwa kwi -25～-15℃, esebenzayo iminyaka emi-2.
Imiyalelo
Le yenye yeendlela eziqhelekileyo zokulungiselela isisombululo se-RNase A.Isenokulungiswa nguezinye iindlela ngokweendlela zemveli elabhoratri okanye uncwadi lwereferensi (njengeukunyibilika ngokuthe ngqo kwi-10 mM Tris-HCl, pH 7.5 okanye isisombululo se-Tris-NaCl)
1. Sebenzisa i-10 mM i-acetate ye-sodium (pH 5.2) ukulungiselela i-10 mg / mL ye-RNase isisombululo sokugcina
2. Ukufudumala kwi-100 ℃ nge-15 min
3. Ukuphola kwiqondo lobushushu begumbi, yongeza i-1/10 yevolumu ye-1 M Tris-HCl (pH 7.4), lungisa i-pH yayo kwi-7.4 (yeumzekelo, yongeza i-500 ml ye-10g / ml isisombululo sokugcina i-RNase 1M Tris-HCL, PH7.4)
4. I-Sub-package kwi -20℃ yokugcinwa komkhenkce, enokuzinza ukuya kwi-2 iminyaka.
[Amanqaku]: Xa ubilisa isisombululo se-RNaseA phantsi kweemeko ezingathathi hlangothi, imvula ye-RNase iya kwenza;Yibilise kwi-pH ephantsi, kwaye ukuba kukho imvula, inokubonwa, enokuthi ibangelwa ubukho bokungcola kweeprotheni.Ukuba intlenga ifunyenwe emva kokubila, ukungcola kunokususwa ngesantya esiphezulu se-centrifugation (13000rpm), kwaye emva koko sipakishwe kancinci ukuze sigcinwe ngumkhenkce.
Ulwazi lwemveliso
| Izithethantonye | Ribonuclease I;i-pancreatic ribonuclease;Ribonuclease 3'-i-pyrimidnooligonucleotidohydrolase;Rnase A;Endoribonulcease I |
| Inombolo yeCAS. | 9001-99-4 |
| Imbonakalo | White lyophilized powder |
| Ubunzima bemolekyuli | ~ 13.7kDa (ulandelelwano lwe-amino acid) |
| Ph Ixabiso | 7.6 (uluhlu lomsebenzi 6-10) |
| Ubushushu obufanelekileyo | 60℃ (Uluhlu lomsebenzi 15-70℃) |
| I-arhente esebenzayo | Na2+.K+ |
| Isithinteli | I-Rnase Inhibitor |
| Indlela yokungasebenzi | Ayinakusebenza ngokufudumeza, kucetyiswa ukuba kusetyenziswe ikholamu ye-centrifuge |
| Imvelaphi | Inkomo |
| Ukunyibilika | Inyibilika emanzini (10mg/ml) |
| Ilahleko ngokomileyo | ≤5.0% |
| Umsebenzi we-enzyme | ≥60 iiyunithi zeKunitz/mg |
| Indawo ye-isoelectric | 9.6 |
Amanqaku
Ukhuseleko kunye nempilo yakho, nceda unxibe iidyasi zaselebhu kunye neeglavu ezilahlwayo xa usebenza.
