Rnasa A
Ribonuclease A (RNaseA) polypeptide ne mai ɗauri guda ɗaya mai ɗauke da 4 disulfide bonds tare da nauyin kwayoyin kusan 13.7 kDa.RNase A shine endoribonuclease wanda ke lalata RNA mai madauri ɗaya musamman a ragowar C da U.Musamman, cleavage ya gane haɗin phosphodiester da aka kafa ta 5'-ribose na nucleotide da ƙungiyar phosphate akan 3'-ribose na pyrimidine nucleotide na kusa, don haka 2, 3'-Cyclic phosphates suna hydrolyzed zuwa daidai 3. 'nucleoside phosphates (misali pG-pG-pC-pA-pG RNase A ce ta kebe don samar da pG-pG-pCp da A-PG).RNase A shine mafi aiki a cikin tsage RNA mai madauri ɗaya.Shawarar aiki maida hankali ne 1-100μG/ml, jituwa tare da daban-daban dauki tsarin.Za'a iya amfani da ƙarancin ƙarancin gishiri (0-100 mM NaCl) don yanke RNA mai ɗauri ɗaya, RNA mai ɗauri biyu, da sarƙoƙin RNA waɗanda aka kafa ta hanyar haɓakar RNA-DNA.Koyaya, a babban maida hankali na gishiri (≥0.3 M), RNase A kawai yana murkushe RNA mai madauri ɗaya.
Ana amfani da RNase A mafi yawa don cire RNA yayin shirye-shiryen DNA na plasmid ko DNA na genomic. Ko DNA yana aiki ko a'a yayin tsarin shirye-shiryen zai iya tasiri cikin sauƙi.Ana iya amfani da hanyar gargajiya ta tafasa a cikin ruwan wanka don hana ayyukan DNAse.Wannan samfurin baya ƙunshi DNAase da protease, kuma baya buƙatar maganin zafi kafin amfani.Bugu da ƙari, ana iya amfani da wannan samfurin a cikin gwaje-gwajen ilimin halitta kamar nazarin kariyar RNase da nazarin jerin RNA.
Yanayin Ajiya
Ana iya adana samfurin a -25 ～-15 ℃, yana aiki na shekaru 2.
Umarni
Wannan shine ɗayan hanyoyin gama gari don shirya mafita na RNase A.Hakanan za'a iya shirya tawasu hanyoyin bisa ga hanyoyin gargajiya a cikin dakin gwaje-gwaje ko wallafe-wallafe (kamarNarkar da kai tsaye a cikin 10 mM Tris-HCl, pH 7.5 ko maganin Tris-NaCl)
1. Yi amfani da 10 mM sodium acetate (pH 5.2) don shirya 10 mg/mL na RNase A ajiya bayani
2. Dumama a 100 ℃ na 15 min
3. Cool zuwa zafin jiki, ƙara ƙarar 1/10 na 1 M Tris-HCl (pH 7.4), daidaita pH zuwa 7.4 (donmisali, ƙara 500 ml na 10g/ml RNase maganin ajiya 1M Tris-HCL, PH7.4)
4. Sub-cushe a -20 ℃ don daskararre ajiya, wanda zai iya zama barga har zuwa shekaru 2.
[Bayanai]: Lokacin tafasa RNaseA bayani a ƙarƙashin yanayin tsaka tsaki, hazo na RNase zai haifar;Tafasa shi a ƙasan pH, kuma idan akwai hazo, ana iya lura da shi, wanda zai iya haifar da kasancewar gurɓataccen furotin.Idan aka sami laka bayan tafasa, za a iya cire ƙazanta ta hanyar centrifugation mai sauri (13000rpm), sannan a yi ƙasa da ƙasa don ajiya mai daskarewa.
Bayanin samfur
| Makamantu | Ribonuclease I;Pancreatic ribonuclease;Ribonuclease 3'-pyrimidnooligonucleotidohydrolase;Rnasa A;Endoribonulcease I |
| CAS No. | 9001-99-4 |
| Bayyanar | Farin lyophilized foda |
| Nauyin kwayoyin halitta | ~ 13.7kDa (jerin amino acid) |
| Ph darajar | 7.6 (Yawan ayyuka 6-10) |
| Dace Zazzabi | 60 ℃ (Ayyukan aiki 15-70 ℃) |
| Wakili mai kunnawa | Na2+.K+ |
| Mai hanawa | Mai hana Rnase |
| Hanyar rashin kunnawa | Ba za a iya kunnawa ta dumama ba, ana ba da shawarar yin amfani da ginshiƙin centrifuge |
| Asalin | Bovine |
| Solubility | Mai narkewa a cikin ruwa (10mg/ml) |
| Asara a bushe | ≤5.0% |
| Enzyme aiki | ≥60 Kunitz raka'a/mg |
| Isoelectric batu | 9.6 |
Bayanan kula
Don amincin ku da lafiyar ku, da fatan za a sa riguna na lab da safofin hannu masu zubarwa don aiki.
