Ultra Nuclease GMP-papa
Helu popoki: HC2016A
Hōʻike ʻia a hoʻomaʻemaʻe ʻia ka UltraNuclease GMP-grade ma Escherichia coli (E.coli) e ka genetically engineered a hoʻomākaukau ʻia ma lalo o nā kaiapuni GMP.Hiki iā ia ke hōʻemi i ka viscosity o ka cell supernatant a me ka cell lysate i ka noiʻi ʻepekema, hoʻonui i ka pono hoʻomaʻemaʻe protein a hoʻonui i ka noiʻi hana protein.Hiki i ka huahana ke hōʻemi i nā koena nucleic acid host i ka pg-grade, e hoʻomaikaʻi i ka hana a me ka palekana o nā huahana olaola o nā noi e pili ana i ka hoʻomaʻemaʻe ʻana i nā maʻi maʻi, ka hana ʻana i nā maʻi, a me ka hana ʻana i nā lāʻau lapaʻau / polysaccharide.Ma waho aʻe o kēia, hiki ke hoʻopili ʻia ka huahana e pale i ka clumping o ke kanaka peripheral blood mononuclear cell (PBMC) i ka cell therapy a me ka hoʻomohala ʻana i ka maʻi.
Hāʻawi ʻia ʻo UltraNuclease ma ke ʻano o kahi reagent sterilized, eluted i ka buffer (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), me ke ʻano o kahi wai kala ʻole, ʻikea.Hana ʻia kēia huahana e nā koi kaʻina GMP a hāʻawi ʻia i kahi ʻano wai.
Nā ʻāpana
UltraNuclease GMP-papa (250 U/μL)
Nā Kūlana Waihona
Hoʻouna ʻia ka huahana me ka hau maloʻo a hiki ke mālama ʻia ma -25 ℃ ~ -15 ° C no ʻelua makahiki.
Inā wehe ʻia ka huahana a mālama ʻia ma 4 ℃ no hoʻokahi pule, manaʻo mākou e kānanaka huahana no ka pale ʻana i ka microbial contamination.
Nā kikoʻī
| Hoʻokipa Hōʻike | Recombinant E. coli me UltraNuclease gene |
| Kaumaha Molekala | 26.5 kDa |
| wahi soelectric | 6.85 |
| Maemae | ≥99% (SDS-PAGE) |
| Hoʻopaʻa mālama | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Wehewehe Unite | ʻO ka wehewehe ʻana o hoʻokahi ʻāpana Hana (U) ka nui o ka enzyme i hoʻohana ʻiahoʻololi i ka waiwai absorption o ΔA260 e 1.0 i loko o 30 minuke i 2. 625 mLʻōnaehana hopena ma 37 ℃ me ka pH o 8.0 (e like me ka hoʻopiha piha ʻana o37 μg salmon sperm DNA i loko o nā oligonucleotides). |
Nā kuhikuhi
1. Laʻana ʻOhi
Nā pūnaewele pili: e wehe i ke kikowaena, holoi i nā keena me ka PBS, a wehe i ka supernatant.
Nā pūnaewele hoʻokuʻu: e hōʻiliʻili i nā pūnaewele ma ka centrifugation, holoi i nā pūnaewele me PBS, centrifuge ma 6,000rpm no 10 min, e ohi i ka pellet.
Escherichia coli: hōʻiliʻili i ka maʻi bacteria ma ka centrifugation, holoi hoʻokahi me PBS, centrifuge ma 8,000rpm no 5 min, a e ohi i ka pellet.
2. Laʻana Lapaʻau
E mālama i nā pellet cell i hōʻiliʻili ʻia me ka lysis buffer ma ka ratio o ka nuipaʻa (g) i ka leo (mL) 1: (10-20), a i ʻole ma ke ʻano mechanical a i ʻole kemika ma ka hau a i ʻole ma ka lumi wela (1g o ka cell pellet aia ma kahi o
109 pūnaewele).
3. Lapaʻau Enzyme
Hoʻohui i ka 1-5mM MgCl i ka ʻōnaehana pane a hoʻoponopono i ka pH i 8-9.
Hoʻohui i ka UltraNuclease e like me ka ratio o 250 Units e ʻeli i 1 g o nā pellet cell, incubate ma 37 ℃ no nā minuke 30.E ʻoluʻolu e nānā i ka palapala "Manawa Manaʻo Manaʻo" e koho i kalōʻihi o ka mālama ʻana.
4. Kahua ʻOhi
Centrifuge ma 12,000 rpm no 30 mau minuke a hōʻiliʻili i ka supernatant.
'Ōlelo Aʻo: Inā ʻakika a alkaline ka hoʻonā, a i ʻole i loko o ka paʻakai kiʻekiʻe, nā mea holoi, a i ʻolenā denaturants, e ʻoluʻolu e hoʻonui i ka nui o ka enzyme a i ʻole e hoʻolōʻihi i ka manawa lapaʻau e like me ia.
Manaʻo ʻia ka hopena hopenaons
| ʻĀpana | Kūlana ʻoi loa | Kūlana kūpono |
| Mg²+Kakau | 1-5 mM | 1-10 mM |
| pH | 8-9 | 6-10 |
| Mahana | 37 ℃ | 0-42 ℃ |
| DTT Hoʻopaʻa | 0-100 mM | >0 mM |
| ʻO Mercaptoethanol Concentration | 0-100 mM | >0 mM |
| Monovalent Cation Concentration | 0-20 mM | 0-150 mM |
| Phosphate lon Concentration | 0-10 mM | 0-100 mM |
Paipai ʻia Pane Manawa (37℃, 2 mM Mg²+, pH 8.0)
| Ka nui o ka UltraNuclease | Manawa Manaʻo |
| 0.25 U/mL | > 10 hola |
| 2.5 U/mL | >4h |
| 25 U/mL | 30 min |
Nā memo:
E ʻoluʻolu e ʻaʻahu i ka PPE pono, e like me ka lole lab a me nā mīkina lima, e hōʻoia i kou olakino a palekana!
