Ultra Nuclease GMP-grade
Nambala ya mphaka: HC2016A
UltraNuclease GMP-grade imasonyezedwa ndikuyeretsedwa mu Escherichia coli (E.coli) ndi majini opangidwa ndi kukonzedwa pansi pa malo a GMP.Itha kuchepetsa kukhuthala kwa ma cell supernatant ndi cell lysate mu kafukufuku wasayansi, kuonjezera kuyeretsa kwa mapuloteni komanso kupititsa patsogolo kafukufuku wamapuloteni.Mankhwalawa amathanso kuchepetsa zotsalira za nucleic acid kukhala pg-grade, kukonza magwiridwe antchito ndi chitetezo chazinthu zachilengedwe zomwe zimagwiritsidwa ntchito kuphatikiza kuyeretsa ma virus, kupanga katemera, ndi kupanga mapuloteni / polysaccharide mankhwala.Kupatula apo, mankhwalawa angagwiritsidwenso ntchito poletsa kuphatikizika kwa maselo amtundu wamagazi amtundu wa mononuclear (PBMC) mu cell therapy ndi chitukuko cha katemera.
UltraNuclease imaperekedwa mu mawonekedwe a sterilized reagent, eluted mu buffer (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), yowoneka ngati madzi opanda mtundu, owonekera.Izi zimapangidwa ndi zofunikira za GMP ndipo zimaperekedwa mu mawonekedwe amadzimadzi.
Zigawo
UltraNuclease GMP-grade (250 U/μL)
Zosungirako
Mankhwalawa amatumizidwa ndi ayezi wouma ndipo amatha kusungidwa pa -25 ℃ ~ -15 ° C kwa zaka ziwiri.
Ngati malonda atsegulidwa ndipo asungidwa pa 4 ℃ kwa nthawi yopitilira sabata, timalimbikitsa kusefamankhwala kuteteza tizilombo toyambitsa matenda.
Zofotokozera
| Expression Host | Recombinant E. coli yokhala ndi jini ya UltraNuclease |
| Kulemera kwa Maselo | 26.5 kda |
| soelectric point | 6.85 |
| Chiyero | ≥99% (SDS-PAGE) |
| Kusungirako Buffer | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Tanthauzo la Chigawo | Tanthauzo la gawo limodzi la Ntchito (U) ndi kuchuluka kwa enzyme yomwe imagwiritsidwa ntchitosinthani mayamwidwe a ΔA260 ndi 1.0 mu mphindi 30 mu 2. 625 mLkachitidwe ka 37 ℃ ndi pH ya 8.0 (yofanana ndi chimbudzi chathunthu cha37 μg umuna wa umuna wa DNA kukhala oligonucleotides). |
Malangizo
1. Chitsanzo Zosonkhanitsa
Maselo omatira: chotsani sing'anga, sambani ma cell ndi PBS, ndikuchotsani zamphamvu.
Maselo oyimitsidwa: sonkhanitsani maselo ndi centrifugation, sambani maselo ndi PBS, centrifuge pa 6,000rpm kwa mphindi 10, sonkhanitsani pellet.
Escherichia coli: sonkhanitsani mabakiteriya ndi centrifugation, sambani kamodzi ndi PBS, centrifuge pa 8,000rpm kwa mphindi 5, ndikusonkhanitsa pellet.
2. Zitsanzo za Chithandizo
Chitani ma cell osonkhanitsidwa ndi lysis buffer pa chiyerekezo cha mass(g) kwa voliyumu(mL)1:(10-20), kapena ndi makina kapena njira zama mankhwala pa ayezi kapena kutentha kwa firiji (1g ya cell pellets ili ndi pafupifupi
109 maselo).
3. Chithandizo cha enzyme
Onjezani 1-5mM MgCl ku machitidwe ndikusintha pH kukhala 8-9.
Onjezani UltraNuclease molingana ndi chiŵerengero cha 250 Units kuti mugaye 1 g ya ma cell a pellets, muyimire pa 37 ℃ kwa mphindi zopitirira 30.Chonde onani fomu ya "Recommended Reaction Time" kuti musankhenthawi ya chithandizo.
4. Wopambana Zosonkhanitsa
Centrifuge pa 12,000 rpm kwa mphindi 30 ndikusonkhanitsa supernatant.
Zindikirani: Ngati yankho lili acidic kapena lamchere, kapena lili ndi mchere wambiri, zotsukira, kapenama denaturants, chonde onjezerani mlingo wa enzyme kapena onjezerani nthawi ya chithandizo moyenerera.
Analimbikitsa reaction conditionse
| Parameter | Mulingo Wabwino Kwambiri | Effective Conditior |
| Mg²+Concentration | 1-5 mm | 1-10 mm |
| pH | 8-9 | 6-10 |
| Kutentha | 37 ℃ | 0-42 ℃ |
| Malingaliro a kampani DTT | 0-100 mm | >0 mM |
| Kukhazikika kwa Mercaptoethanol | 0-100 mm | >0 mM |
| Monovalent Cation Concentration | 0-20 mm | 0-150 mm |
| Phosphate Lon Concentration | 0-10 mm | 0-100 mm |
Analimbikitsa Zomwe anachita Nthawi (37 ℃, 2 mM Mg²+, pH 8.0)
| UltraNuclease Money (Final Concentration) | Nthawi Yochitira |
| 0.25 U/mL | >10h |
| 2.5 U/mL | > 4h |
| 25 U/mL | 30 min |
Ndemanga:
Chonde valani PPE yofunikira, malaya a labu ndi magolovesi, kuti mutsimikizire thanzi lanu ndi chitetezo!
