Rnase A
ʻO Ribonuclease A (RNaseA) kahi polypeptide i hoʻopaʻa ʻia me 4 mau paʻa disulfide me ke kaumaha molekala ma kahi o 13.7 kDa.ʻO RNase A kahi endoribonuclease e hoʻohaʻahaʻa pono i ka RNA kaula hoʻokahi ma nā koena C a me U.ʻO ke kikoʻī, ʻike ka cleavage i ka paʻa phosphodiester i hoʻokumu ʻia e ka 5'-ribose o kahi nucleotide a me ka hui phosphate ma ka 3'-ribose o ka nucleotide pyrimidine e pili ana, no laila ua hydrolyzed ka 2, 3'-Cyclic phosphates i ka 3 pili. 'nucleoside phosphates (e laʻa me pG-pG-pC-pA-pG i hoʻokaʻawale ʻia e RNase A e hoʻohua i ka pG-pG-pCp a me A-PG).ʻO RNase A ka mea ikaika loa i ka hoʻokaʻawale ʻana i ka RNA kaula hoʻokahi.ʻO 1-100μG/mL ka manaʻo hana i manaʻo ʻia, kūpono me nā ʻōnaehana like ʻole.Hiki ke hoʻohana ʻia ka paʻakai haʻahaʻa (0-100 mM NaCl) no ka ʻoki ʻana i ka RNA kaula hoʻokahi, ka RNA pālua, a me nā kaulahao RNA i hana ʻia e ka RNA-DNA hybridization.Eia nō naʻe, ma ka paʻakai kiʻekiʻe o ka paʻakai (≥0.3 M), ʻo RNase A wale nō e hoʻokaʻawale i ka RNA kaula hoʻokahi.
Hoʻohana pinepine ʻia ʻo RNase A no ka wehe ʻana i ka RNA i ka wā o ka hoʻomākaukau ʻana i ka plasmid DNA a i ʻole genomic DNA. ʻO ka hana ʻana a i ʻole ʻole ʻo DNase i ka wā o ka hoʻomākaukau ʻana hiki ke hoʻopili maʻalahi i ka hopena.Hiki ke hoʻohana ʻia ke ʻano kuʻuna o ka paila ʻana i ka ʻauʻau wai e hoʻopau i ka hana DNase.ʻAʻole loaʻa i kēia huahana ka DNase a me ka protease, a ʻaʻole koi i ka mālama wela ma mua o ka hoʻohana ʻana.Eia kekahi, hiki ke hoʻohana ʻia kēia huahana i nā hoʻokolohua molecular biology e like me ka nānā ʻana i ka pale o RNase a me ka nānā ʻana i ke kaʻina RNA.
Nā Kūlana Waihona
Hiki ke mālama ʻia ka huahana ma -25～-15 ℃, kūpono no 2 mau makahiki.
Nā kuhikuhi
ʻO kēia kekahi o nā ala maʻamau no ka hoʻomākaukau ʻana i ka hoʻonā mālama mālama RNase A.Hiki ke hoʻomākaukau ʻia enā ʻano hana ʻē aʻe e like me nā ʻano kuʻuna i loko o ka hale hana a i ʻole nā palapala kuhikuhi (e like mehoʻoheheʻe pololei ʻia i 10 mM Tris-HCl, pH 7.5 a i ʻole Tris-NaCl solution)
1. E hoʻohana i ka 10 mM sodium acetate (pH 5.2) no ka hoʻomākaukau ʻana i ka 10 mg/mL o RNase A hoʻonā mālama.
2. Hoʻomaʻamaʻa ma 100 ℃ no 15 min
3. Hoʻolili i ka lumi wela, hoʻohui i ka 1/10 volume o 1 M Tris-HCl (pH 7.4), hoʻoponopono i kona pH i 7.4 (no ka mea.laʻana, hoʻohui i 500 ml o 10g/ml RNase waihoʻoluʻu 1M Tris-HCL, PH7.4)
4. Sub-packaged ma -20 ℃ no ka mālama ʻana i ka hau, hiki ke paʻa a hiki i 2 mau makahiki.
[Nā memo]: I ka hoʻolapalapa ʻana i ka solution RNaseA ma lalo o nā kūlana kū ʻole, e hoʻokumu ʻia ka ua o RNase;E hoʻolapalapa iā ia ma kahi pH haʻahaʻa, a inā he ua, hiki ke ʻike ʻia, hiki ke kumu ma muli o ka loaʻa ʻana o nā haumia protein.Inā loaʻa ka sediment ma hope o ka hoʻolapalapa ʻana, hiki ke hoʻoneʻe ʻia nā mea haumia e ka centrifugation kiʻekiʻe (13000rpm), a laila hoʻopiha piha ʻia no ka mālama hau.
ʻIke huahana
| Nā huaʻōlelo like | Ribonuclease I;Pancreatic ribonuclease;Ribonuclease 3'-pyrimidnooligonucleotidohydrolase;Rnase A;ʻO Endoribonulcease I |
| CAS No. | 9001-99-4 |
| Ka nana aku | ʻO ka pauka lyophilized keʻokeʻo |
| Kaumaha molekula | ~ 13.7kDa (ke kaʻina amino acid) |
| Ph Waiwai | 7.6 (nui hana 6-10) |
| ʻO ka Mahana kūpono | 60 ℃ (Mahele hana 15-70 ℃) |
| Mea ho'āla | Na2+.K+ |
| Kāohi | Rnase Kāohi |
| ʻano hana ʻole | ʻAʻole hiki ke hoʻopau ʻia e ka hoʻomehana, ʻōlelo ʻia e hoʻohana i ke kolamu centrifuge |
| Ke kumu | Bovine |
| Ka hikiwawe | Hiki i ka wai (10mg/ml) |
| Nalo ma kahi maloʻo | ≤5.0% |
| Ka hana enzyme | ≥60 Kunitz wae/mg |
| kiko isoelectric | 9.6 |
Nā memo
No kou palekana a me kou olakino, e ʻoluʻolu e ʻaʻahu i nā ʻaʻahu lab a me nā mīkina lima hoʻopau no ka hana.
