IZON qEV1 Columns User Guide

qEVORIGINAL Gen 2 Magawo
Buku Lophunzitsira
KWA qEVORIGINAL GEN 2 COLUMNS (35 nm & 70 nm)

Chitsogozo choyambira mwachanguchi chimapereka malangizo ogwiritsira ntchito.
Kuti mumve zambiri, mutha kutsitsa laibulale yonse ya qEV User Manuals ndi zinthu zina kuchokera patsamba lothandizira la Izon pa. support.izon.com

Safety Data Sheets akupezeka pa support.izon.com/safety-datasheets

NTCHITO YOTSATIRA

Mizati ya qEV imapatula ma vesicles owonjezera kuchokera ku biological samples ndipo ali ndi tchipisi ta RFID kuti mugwiritse ntchito ndi Automatic Fraction Collector (AFC).
Ma chips awa sangakhudze kugwiritsa ntchito pamanja.

Magawo a qEV amapangidwa kuti azigwiritsidwa ntchito m'ma laboratories ofufuza ndi akatswiri kuti agwiritse ntchito kafukufuku kokha. Magawo a qEV sali ndi cholinga chowunikira matenda ndipo sayenera kugwiritsidwa ntchito popanga chisankho chamankhwala.

MALANGIZO OGWIRA NTCHITO

1. Centrifuge samppang'ono asanakweze ndime kuti achotse ma cell ndi zinyalala zazikulu zama cell. Poyamba centrifuge pa 1,500 xg kwa mphindi 10 kuchotsa maselo aliwonse ndi tinthu tating'onoting'ono. Rencentrifuge the supernatant pa 10,000 xg kwa mphindi 10.
2. Kwa magulu akuluakulu samples, n'zotheka kuika maganizo a sample musanayike pagawo la qEV. Izi sizikugwira ntchito ku seramu ndi plasmaamples, omwe ali ndi mapuloteni ochuluka kwambiri. Izon amalimbikitsa kugwiritsa ntchito zida za Merck Millipore, ndi zosefera za Amicon® Ultra Centrifugal.
3. Izon imalimbikitsa kugwiritsa ntchito mizati kamodzi ngati mukufuna kusanthula ma vesicles a nucleic acid.
4. Onetsetsani kuti sample buffer ili pa kutentha komweko monga gawoli (makamaka pakati pa 18-24 ˚C).
5. Gwiritsani ntchito buffer yatsopano (0.22 µm) kuti mupewe kuipitsidwa.

KULETSA MALANGIZO

qEV1 COLUMN ZINTHU

Sampndi Load Volume	≤ 1 mL
Column Volume	13.5 mL
Buffer Volume	4.0 mL
Kukula Kwagawo Loyenera	0.7 mL
Buffer yofunikira pa sample zosonkhanitsira	10.0 mL

KULINGANA

1. Gwirizanitsani ndime ndi sampChosungiracho chiyenera kukhala mkati mwa kutentha kwa 18-24 ˚C. Osachotsa zipewa mpaka kutentha kwa ntchito kufikire.
2. Chotsani mosamala chipewa chapamwamba chokha.
3. Ikani mzatiwo mowongoka pa choyikapo chokonzekera kugwiritsidwa ntchito. Ma qEV Racks ndi Automatic Fraction Collectors akupezeka ku Izon Science.
4. Ikani nkhokwe ngati ilipo, ndipo onjezerani buffer.

KUPULUKA KWA NTHAWI

1. Chotsani kapu yapansi ndikulola kuti buffer iyambe kudutsa ndime.
2. Yambani ndime ndi gawo limodzi la buffer. Ngati mapulogalamu anu akumunsi akhudzidwa ndi sodium azide, tsitsani ndi ma voliyumu awiri a bafa. Ngati chotchinga chotchinga china kupatula PBS ndichoti chigwiritsidwe ntchito, gwirizanitsani gawoli ndi magawo atatu a bokosi latsopanolo. Mzerewu udzasiya kuyenda pokhapokha pamene ma buffers onse alowa mu frit yotsegula.

MANUAL SAMPLE KUSONYEZA

1. Sefa kapena centrifuge biological sample kuchotsa zinthu zazikulu.
   Onani zomwe zaperekedwa pamwambapa.
2. Chosungiracho chikasiya kuyenderera muzanja kuti isasunthike, kwezani ma centrifuged okonzekaamplembani voliyumu ku frit yotsitsa.
   Pewani kuyimitsa mzati pakuyenda kwa nthawi yayitali kuti muwonetsetse kulekanitsidwa kolondola kwa EV.
3. Nthawi yomweyo yambani kutolera buffer volume1 (izi zikuphatikiza voliyumu yomwe idachotsedwa ndikutsitsa sample).
4. Lolani sample kuthamangira mu column. Mzere udzasiya kuyenda pamene onse a samples alowa mu loading frit.
5. Kwezani mosungiramo / gawoli ndi buffer ndikupitiliza kusonkhanitsa voliyumu ya bafa.
   Kuti mutenge ma voliyumu olondola, ingokwezani voliyumu yomwe ikufunika pamwamba pa ndimeyo, dikirani kuti voliyumu idutse mpaka kutuluka kutayima, ndikubwereza. 
6. Voliyumu ya buffer ikasonkhanitsidwa, pitilizani kusonkhanitsa Purified Collection Volume (PCV)2. Onani Zithunzi 1 ndi 2.

COLUMN FLUSH NDI KUSINTHA

1. Ma voliyumu omwe mukufuna atasonkhanitsidwa, tsitsani gawoli ndi 8.5 mL ya 0.5 M Sodium Hydroxide (NaOH) ndikutsatiridwa ndi 17 mL ya buffer musanalowetsenso s.ample.
2. Ngati mukusunga mzati kuti mudzagwiritse ntchito mtsogolo, yambani ndi bafa yokhala ndi bacteriostatic agent (monga 0.05 % w/v sodium azide), kapena 20% ethanol.
3. Mipingo imatha kusungidwa kutentha kwa chipinda mukatha kugwiritsidwa ntchito, malinga ngati yatsukidwa molingana ndi malangizo omwe ali pamwambapa. Ngati njira zoyenera sizikupezeka ndiye kuti mizati ikhoza kusungidwa pa 4-8 ˚C mukatha kugwiritsa ntchito.

¹Voliyumu ya buffer - kuchuluka kwa bafa komwe kumatuluka pamndandanda wa tinthu tating'ono ta chidwi.
²Purified Collection Volume (PCV) - voliyumu yoyeretsedwa ndi gawo lomwe lili ndi tinthu tating'ono ta chidwi.
Onani buku lathunthu la ogwiritsa ntchito kuti mudziwe zambiri posankha PCV yoyenera.

Chithunzi 1. Wodziwika bwino wa elutionfile kwa qEV1 35 nm ndime yokhala ndi 1.0 mL yamadzi am'magazi amunthu yodzaza; Mapuloteni amatuluka mochedwa kuposa ma vesicles. Kuchuluka kwa vesicle kunayesedwa pogwiritsa ntchito Exoid ndi mapuloteni ndi kusanthula kwa BCA.

EVs ndi tinthu tating'ono tofanana
Mulingo wa mapuloteni

Chithunzi 2. Kuyerekeza kwa mapuloteni ndi EV elution profiles pakati pa qEV1 35 nm ndi qEV1 70 nm. Mizati yofotokozedwa ndi mfundo zongoyerekeza zotengera ma s ophatikizidwaample miyeso.

qEV/35 EVs ndi tinthu tating'ono tofanana
qEV/35 mulingo wa protein
qEV/70 EVs ndi tinthu tating'ono tofanana
qEV/70 mulingo wa protein

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Zolemba / Zothandizira

Zithunzi za IZON qEV1 [pdf] Wogwiritsa Ntchito qEV1 Columns, qEV1, Columns

Zothandizira

• Izon Science Support Center
• Dzipatulani & Muyese Zowonjezera Zowonjezera & Nanoparticles
• Manual wosuta