Ultra Nuclease GMP-giredhi
Katsi Nhamba: HC2016A
UltraNuclease GMP-giredhi inoratidzwa nekucheneswa muEscherichia coli (E.coli) nemajini enjiniya uye yakagadzirirwa pasi penzvimbo dzeGMP.Inogona kuderedza viscosity yesero supernatant uye sero lysate mukutsvagisa kwesainzi, kuwedzera mapuroteni ekunatsa kushanda zvakanaka uye kuwedzera mapuroteni anoshanda ekutsvaga.Chigadzirwa chacho chinogona zvakare kudzikisa host nucleic acid zvakasara kusvika pg-giredhi, kuvandudza mashandiro uye kuchengetedzeka kwezvinhu zvipenyu zvekushandisa zvinosanganisira kucheneswa kwehutachiona, kugadzira vaccine, uye mapuroteni/polysaccharide kugadzirwa kwemishonga.Kunze kwezvo, chigadzirwa chinogona zvakare kuiswa kudzivirira kuunganidzwa kweropa remunhu mononuclear masero (PBMC) musero kurapa uye kuvandudza vaccine.
UltraNuclease inopiwa muchimiro cheajenti yakaomeswa, yakaburitswa mubuffer (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), ine chitarisiko chemvura isina ruvara, inoonekera.Ichi chigadzirwa chinogadzirwa neGMP process zvinodiwa uye chinopihwa mune yemvura fomu.
Zvikamu
UltraNuclease GMP-giredhi (250 U/μL)
Storage Conditions
Chigadzirwa chacho chinotumirwa nechando chakaoma uye chinogona kuchengetwa pa -25 ℃ ~ -15 ° C kwemakore maviri.
Kana chigadzirwa chakavhurwa uye chachengetwa pa4 ℃ kweanopfuura vhiki, tinokurudzira kusefachigadzirwa kudzivirira microbial kusvibiswa.
Zvinotsanangurwa
| Expression Host | Recombinant E. coli ine UltraNuclease gene |
| Molecular Weight | 26.5 kDa |
| soelectric point | 6.85 |
| Kuchena | ≥99% (SDS-PEJI) |
| Kuchengetedza Buffer | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Chikamu Tsanangudzo | Tsanangudzo yeimwe Chiitiko unit (U) huwandu hwee enzyme inoshandiswashandura kukosha kwekunyura kweΔA260 ne1.0 mumaminitsi makumi matatu mune 2. 625 mLreaction system pa37 ℃ ine pH ye8.0 (yakaenzana nekupedza kugaya kwe37 μg salmon urume DNA mune oligonucleotides). |
Mirayiridzo
1. Muenzaniso Collection
Adherent masero: bvisa yepakati, geza maseru nePBS, uye bvisa iyo supernatant.
Maseru ekusimudzwa: unganidza maseru ne centrifugation, geza maseru nePBS, centrifuge pa6,000.rpm kwemaminetsi gumi, unganidza pellet.
Escherichia coli: unganidza bhakitiriya necentrifugation, geza kamwe chete nePBS, centrifuge pa8,000.rpm ye5 min, uye tora pellet.
2. Muenzaniso Kurapa
Bata maseru akaunganidzwa ne lysis buffer pachiyero chehuremu(g) kusvika kuvhoriyamu(mL)1:(10-20), kana nemakanika kana makemikari nzira paaizi kana patembiricha yekamuri (1g yesero pellet rinenge
109 masero).
3. Enzyme Kurapa
Wedzera 1-5mM MgCl kune reaction system uye gadzirisa pH kusvika 8-9.
Wedzerai UltraNuclease maererano nereshiyo ye250 Units kuti mugaye 1 g yemacell pellets, incubate pa37℃ kweanopfuura maminetsi makumi matatu.Ndokumbira utarise kune "Yakakurudzirwa Rection Nguva" fomu kuti usarudze iyonguva yekurapa.
4. Supernatant Collection
Centrifuge pa12,000 rpm kwemaminetsi makumi matatu uye tora iyo supernatant.
Ongorora: Kana mhinduro yacho iine acidic kana alkaline, kana ine yakanyanya kuwanda yemunyu, detergents, kanadenaturants, ndapota wedzera iyo enzyme dosage kana kuwedzera nguva yekurapa zvinoenderana.
Recommended reaction conditions
| Parameter | Optimal Condition | Effective Conditior |
| Mg²+Concentration | 1-5 mM | 1-10 mm |
| pH | 8-9 | 6-10 |
| Tembiricha | 37℃ | 0-42 ℃ |
| DTT Concentration | 0-100 mm | >0 mM |
| 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 |
Recommended Reaction Nguva (37℃, 2 mM Mg²+, pH 8.0)
| UltraNuclease Amount (Final Concentration) | Reaction Time |
| 0.25 U/mL | >10 h |
| 2.5 U/mL | >4h |
| 25 U/mL | 30 min |
Notes:
Ndokumbira upfeke PPE inodiwa, jasi rakadaro uye magurovhosi, kuve nechokwadi chehutano hwako uye kuchengetedzeka!
