Pichia liwiro la phofi
Pichia liwiro la phofi
✓ Pichia Abusa - Palibe Mtanda - Kutaya Out / Hcps
✓ Wokonzeka - Gwiritsani - Palibe zokopa zomwe zimafunikira
Kugwiritsa Ntchito
Kityi idapangidwira kupezeka kwa ochulukitsa kwa protein (HCP) ku biopharautical zinthu zochokera ku Pichia litoris Zovuta (E.g., GS115, x33), kugwiritsa ntchito imodzi - cholumikizira enzyme - Chimodzimodzi Kufufuza kokha. Osati kuti zizindikiritso.
Mfundo Yodziwikiratu
Chingwe chimagwiritsa ntchito a sangweji Elisa Njira:
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Chokutila: Zitsime za Microplate ndi Pres Pre - zokutidwa ndi ma antibodies a nkhosa mokwanira Pichia litoris HCPS.
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Makulitsidwe: Samclec / Acaribratoors amawonjezeredwa, kenako Hrkp - Ophatikizidwa ndi Nkhosa a Polyclol Anti -Pichia litoris Ma antibodies hcp.
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Kufufuza: Mukasambitsidwa, TMB gawo la TMB limawonjezeredwa chifukwa cha chitukuko cha utoto. Zomwe zimachitika, ndipo kuyamwa kumayesedwa 450 nm.
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Ndalama: HCP ndende imawerengeredwa kuchokera ku curve wokhazikika (wopezeredwa α hcp ndende).
Mawonekedwe Ofunika
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Palibe zokopa zomwe zimafunikira (kutsimikizika kokha komwe pakufunika).
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Protocol yokhazikika: Masitepe ochepa, zotsatira zachangu (<2 maola).
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Kuperewera kwakukulu komanso kudalirika: Okhazikika Pichia litoris HCPS.
Zolemba zaukadaulo
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Zitsanzo Zogwirizana: Chikhalidwe cha Cell, zikhalidwe zoyeretsa, zoyeretsa biologics.
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Mitundu yamphamvu: 1-100 ng / ml (monga).
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Zowongolera Zowongolera: Zoyenera kusintha mapangidwe ndi gmp Loti Kuyesa.
Njira yogwiritsira ntchito1. Kufanana
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Bweretsani zinthu zonse kutentha kwa chipinda (≥2 mphindi).
2. Zowonjezera / zowonjezera-
Onjezani zitsanzo / calibrators: 100 μl / chabwino
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Onjezani 1x Enzyme - Wophatikizidwa ndi Antibody: 100 μl / chabwino (onani nthano ①)
3. Makutu-
Tratubera ku RT, Otetezedwa ku kuwala, 600 rpm kwa 3 h (onani nthano ③)
4. Kusamba-
Sambani 1: 300 μl / chabwino 1x buffer (malo ofukula a mbale)
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Sambani 2 - 5: 300 μl / chabwino 1x Buffer, 5
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Pat Dulani: Pambuyo pa kusamba komaliza, kulowa pa pepala loyamwa
5. TMB Gawo-
Onjezani TMB: 100 μl / chabwino
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Kulitsa: 10 min ku RT (kutetezedwa ku kuwala)
6. Lekani Kuchita & Kuwerenga-
Onjezani yankho: 50 μl / chabwino
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Ima: 5 min
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Werenga: Od pa 450 nm (Reference 620 nm)
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| Chipangizo | Zogulitsa Ayi. | Chifanizo | Kaonekeswe |
|---|---|---|---|
| Pre - Olimbana - Pichia HCP Microphete | Pn015 | 8 Wells × 12 | Pres - Ouzidwa ndi Nkhosa Polyclol Anti -Pichia litoris Ma antibodies hcp. Wosindikizidwa m'matumba a zojambula ndi decoccant. Sungani otetezedwa ku kuwala. |
| Pichia hcp calibrator | PNB015 | 2 Mbale | Wadzi ufa. Ngonja ndi 500 μl calibrator (PNC002). Lolani 5-10 min kuti musungunuke (yankho liyenera kukhala lowonekeratu). Sungani otetezedwa ku kuwala. |
| Kalibitor wamphamvu | PNC002 | 1.5 ml × 1 chubu | Yankho lomveka bwino la Pichia Hcp Kalibrator. |
| Kukakamiza | Pne006 | 25 ml × 2 mabotolo | Chifukwa chosakanikirana, zitsanzo, ndi ma enzyme - Wopanga antibody. Vomerezani vuto lokwanira la zitsanzo zatsopano. |
| Sambani buffer (10 × yikani) | Pnf001 | 25 ml × 2 mabotolo | Kutsuka mbale. Makhiristo amatha kupanga kutentha pang'ono - kusungunuka pa 37 ° C asanagwiritse ntchito. Kuchepetsa 10 × ndi madzi okwanira. |
| HRP - Wophatikizidwa anti - Pichia HCP antibody (100 ×) | Pnn009 | 120 μL × 1 chubu | Nkhosa za polyclonal antibody. Kuchepetsa 100 × popewa musanagwiritse ntchito. Sungani otetezedwa ku kuwala. |
| TMB gawo | Pnd004 | 12 ml × 1 botolo | Kufanana ndi kutentha kwa chipinda (≥20 min) musanagwiritse ntchito. Sungani otetezedwa ku kuwala. |
| Siyani yankho | Pni002 | 6 ml × 1 botolo | Ili ndi HCL. Valani zigawenga ndikupewa kulumikizana ndi khungu. |
| Mafilimu | Pnki001 | 3 ma sheet | Kuti kuphimba microphelate kuletsa kuipitsidwa ndi kutuluka. |
Kutumiza Zambiri
Timapereka kunyamula mayendedwe onse pamalamulo onse. Nthawi zambiri, oda yanu ifika mkati mwa 5 - masiku 7 a bizinesi ku United States komanso mkati mwa masiku 10 antchito a mayiko ena. Komabe, chonde dziwani kuti kupembedza kumidzi ingatenge nthawi yayitali.
Nthawi Yotumiza: Maodawo amakonzedwa mkati mwa 1 - masiku atatu antchito. Dongosolo lanu litatumizidwa, mudzalandira imelo yotsimikizira ndi chidziwitso chotsatira.
Chidziwitso chofunikira
Kukonzanso: Kulamula kwaperekedwa, nyumba yathu yosungirako ikusowa nthawi kuti akonze dongosolo lanu. Mudzalandira chidziwitso kamodzi oda yanu yatumizidwa.
Nthawi zoperekera: Nthawi zambiri, phukusi lidzaperekedwa mkati mwa nthawi yomwe ikuchitika. Komabe, tsiku lenileni la zoperekera lingakhudzidwe ndi makonzedwe, nyengo, ndi zinthu zina zakunja. Nthawi yoperekera idzakhala yotalikirapo kuposa masiku onse okhudzana ndi malo ophatikizika kapena zinthu zosinthidwa. Chonde onani chidziwitso chotsatira tsiku lolondola kwambiri.
Nkhani Zotumiza: Ngati mukuwona kuti phukusi lanu silinaperekedwe mkati mwa nthawi yodziwika; Zambiri zotsatila zikuwonetsa kuti phukusi laperekedwa koma simunalandire; kapena phukusi lanu limaphatikizapo zinthu zosowa kapena zolakwika kapena zovuta zina, chonde lemberani kasitomala mkati mwa masiku 7 a tsiku lolipira kuti tithe kuthana ndi mavutowa mwachangu.
Kutsatira kutsatira
Dongosolo lanu litatumizidwa, mudzalandira imelo ndi nambala yolondola komanso ulalo kuti muthe kutumiza.
Mutha kutsatanso dongosolo lanu mwachindunji patsamba lathu podula muakaunti yanu ndikuwona mbiri yanu.
Zoletsa Kutumiza
Chonde lembani adilesi yamsewu mwatsatanetsatane, osati bokosi kapena adilesi yankhondo (apo). Kupanda kutero, tiyenera kugwiritsa ntchito EMS kuti tibweretsenso ena (amayamba pang'onopang'ono kuposa ena, kutenga 1 - miyezi iwiri kapena kupitilira).
Ntchito Zogwirira Ntchito ndi Msonkho
Chonde dziwani kuti ntchito zilizonse zamitundu, misonkho, kapena zolipirira zobwereketsa zomwe zikugulitsidwa ndi udindo wa wogula. Zolipira izi zimasiyana kutengera dzikolo ndipo zimatsimikizika ndi akuluakulu akwanuko am'deralo.
Pogula kuchokera patsamba lathu, mukuvomera kulipira ntchito kapena misonkho yomwe imagwirizana ndi dongosolo lanu. Sitili ndi vuto la kuchedwa chifukwa cha chilolezo cha miyambo.
MALANGIZO OTHANDIZA
Dongosolo lanu litafika pamalo osankhidwa kapena malo operekera, chonde onetsetsani kuti ikutola. Ngati phukusi silinatengedwe mkati mwa nthawi yomwe yasankhidwa, tidzakutumizirani chikumbutso kudzera pa imelo kapena SMS. Komabe, ngati phukusi silikusonkhanitsidwa nthawi yayitali, ndipo kutayika kapena kuwonongeka kulikonse kumachitika chifukwa, wogula azikhala ndi mlandu. Tikukumbutsani mokoma kuti mutole phukusi lanu mwachangu kupewa nkhani zilizonse zomwe zingachitike.
Chidziwitso: Pamene malonda athu amagwera pansi pa gulu lapadera, amabwerera ndi kubweza sikuvomerezedwa.
