PICHIA PRODORIS HCP (Sispotion stilin) Kit
PICHIA PRODORIS HCP (Sispotion stilin) Kit
✓ Pichia Pastis - Ntho e ikhethang - Ha ho na Cross - Boikoetliso ba Cho / HCPS
✓ E itokisitse - ho - Sebelisa - Ha ho na mohlala oa mohlala o hlokahalang
Ts'ebeliso e reriloeng
Kit e etselitsoe ho khetholloa ha palo ea li-protein ea li-spropit tsa li-prosen (HCP) lihlahisoang tsa li-biopharmaceutical li tsoa ho Pichia Pastoris Mathata (e.g.., GS115, X33) Bakeng sa ts'ebeliso ea lipatlisiso feela. Eseng ka sepheo sa ho nka nako.
Molao-motheo oa ho lemoha
Kit e sebelisa a Sandwich Elisa Mokhoa:
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Koae: Lits'ebeletso tsa Microlayplate li pele - e koahetsoe ke li-antibodies tsa linku tse khahlanong le polychal khahlanong le Pichia Pastoris HCPS.
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Incubation: Li-camples / Calibrations li ekelitsoe, lateloa ke HRP - Linku tsa linku tsa polygnal anti -Pichia Pastoris HCP li-antibodies.
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Ho lemohaKamora ho hlatsoa, ho kenyelletsa lekgolo ho eketsoa bakeng sa nts'etsopele ea mmala. Karabelo e emisitsoe, 'me ho kena-kenana ho lekanngoa ho 450 nm.
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QETELA: HCP khatello ea maikutlo e baloa ho tsoa ho li-curve tse tloaelehileng (ho monya α hcp khatello ea maikutlo).
LINTHO TSE HLAHANG
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Ha ho na mohlala oa mohlala o hlokahalang (Ho hlokahala netefatso e hlokahalang).
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E fumane protocol: Mehato e fokolang, liphetho tse potlakileng (
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Phapang e phahameng le ho ts'epahala: E ntlafalitsoe bakeng sa Pichia Pastoris HCPS.
LITLHAKISO TSA BOPHELO
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Disampole tse lumellanang: Li-Supenatants, baphologics ba balolo ba hloekisoang.
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Rannamic: 1-100 NG / ml (e tloaelehileng).
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Alognment ea taolo: E loketse ts'ebetso ea ts'ebetso le tlhahiso ea gmp lotho ho lokolla liteko.
Ts'ebetso ea ts'ebeliso1. Karotination
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Tlisa likarolo tsohle ho mocheso oa kamore (≥20 min).
2. Mohlala oa mohlala / tlatsetso ea antibody-
Kenya sampole / Calibration: 100 μl / hantle
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Kenya 1x enzyme - hlakola antibody: 100 μl / hantle (bona Legend ①)
3. Introbation-
Incubte inst, e sirelelitsoeng ho tloha lebone, 600 rpm bakeng sa 3 h (bona Legend ③)
4. Ho hlatsoa-
Hlatsoa 1: 300 μl / Well 1x buffer (boemo ba poleiti ea vertical)
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Hlatsoa 2 - 5: 300 μl / Well 1x buffer, 5 e hlatsoa
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Pat o omileKa mor'a ho lieha ho hlatsoa, ho hlopha poleiti ho pampiri e monyang
5. TMB Equight nts'etsopele-
Eketsa tmb: 100 μl / hantle
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Ntlafatsa: 10 min ho RT (e sirelelitsoeng ho tloha lebone)
6. Emisa karabelo le ho bala-
Kenya tharollo ea emisa: 50 μl / hantle
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Ema: 5 min
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Bala: Od ka 450 nm (Reference 620 NM)
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| Karolo | Sehlahisoa. | TLHOKOMELISO | Tlhaloso |
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| Pre - Anti - Tachia HCP Microplay | Pna015 | Li-Wells tse 8 × 12 liliba | Pre - koae ka peo ea linku polygnal anti -Pichia Pastoris HCP li-antibodies. E tiisitsoe ka mehala ea foil e nang le lefuba. Lebenkele le sirelelitsoe leseling. |
| Tachia HcP Calibrari | Pnb015 | Li-vielis tse 2 | Phofo ea lyophilized. E pheta-pheta ka calibration ea 500 μl dμl dount (pnc002). Lumella 5-10 mpeng ea ho qhalana (tharollo e lokela ho hlakisa). Lebenkele le sirelelitsoe leseling. |
| Calibration e hlasimollang | Pnc002 | 1.5 ml × 1 tube | Thakallo e hlakileng ea ho pheta taelo ea Pichia HCP. |
| Assan Duient | Pne006 | 25 ml × 2 botlolo | Bakeng sa ho hlakola lik'hamera, sampole, le enzyme - ha etisoa antibody. Netefatsa ntho e phethahetseng ea liksepole tse ncha. |
| Hlatsoa buffer (10 × tsepaminta) | Pnf001 | 25 ml × 2 botlolo | Bakeng sa ho hlatsoa poleiti. Likristale li ka hlaha mochesong o tlase-li qhibilihang ka 37 ° C pele a sebelisoa. Clute 10 × ka metsi. |
| HRP - Hlahloba Anti - Tachia HCP Antibody (100 ×) | Pnn009 | 120 μl × 1 tube | Linku tsa polyclon antibody. Dilute 100 × ka matla a ho sebelisana pele a sebelisoa. Lebenkele le sirelelitsoe leseling. |
| Tmb substrate | Pnd004 | 12 ml × 1 botlolo | Ho lekana ho mocheso oa kamore (≥20 min) pele o sebelisoa. Lebenkele le sirelelitsoe leseling. |
| Emisa tharollo | Pni002 | 6 ml × 1 Bottle | E na le hcl. Roala li-goggles le ho qoba ho ikopanya le letlalo. |
| Ho netefatsa lifilimi | Pnk001 | Lishiti tse 3 | Bakeng sa sekoaelo se akaretsang makala a microplat ho thibela tšilafalo le mouoane. |
Tlhahisoleseling ea ho tsamaisa
Re fana ka lipalangoang tse sephara ho litaelo tsohle. Ka tloaelo, odara ea hau e tla fihla leqepheng la 5 - matsatsi a khoebo a 7 United States le matsatsing a 10 a khoebo bakeng sa linaha tse ling. Leha ho le joalo, ka kopo hlokomela hore ho isoa ho libakeng tsa mahaeng ho ka nka nako e telele.
Nako ea ho tsamaisa: Litaelo li sebetsoa nakong ea 1 - matsatsi a 3 a khoebo. Hang ha odara ea hau e se e rometsoe, u tla fumana lengolo-tsoibila le netefatsang ka tlhaiso-leseling.
Tlhahisoleseling ea Bohlokoa
Ho sebetsa ha taolo: Kamora hore taelo e se e lefiloe, ntlo ea rona ea bophelo e hloka nako ea ho sebetsa odara ea hau. U tla fumana tsebiso hang ha odara ea hau e rometsoe.
Mefuta ea ho tsamaisa: Maemong a mangata, sephutheloana se tla tsamaisoa ka nako e lekantsoeng ea ho fihla. Leha ho le joalo, letsatsi la pele la tlhahiso le ka angoa ke litokisetso tsa lifofane, maemo a leholimo le lintho tse ling tsa kantle. Nako ea ho tsamaisa thepa e tla be e le telele ho feta e tloaelehileng bakeng sa litaelo tse kenyelletsang mokhoa oa pele kapa lintho tse hlophisitsoeng. Ka kopo sheba tlhaiso-leseling ea ho batla bakeng sa letsatsi le nepahetseng ka ho fetisisa.
Litaba tsa ho tsamaisa: Haeba u fumana hore sephutheloana sa hau ha se so lopolloe ka nako e boletsoeng; Tlhahisoleseling ea ho lelekisa e bontša hore sephutheloana se pholositsoe empa ha u e amohetse; Kapa sephutheloana sa hau se kenyelletsa lintho tse sieo kapa tse fosahetseng kapa mathata a mangata, ka kopo ikopanye le ts'ebeletso ea bareki nakong ea matsatsi a 7 e le hore re ka sebetsana le litaba tsena hang-hang.
Ho latela tumello
Hang ha odara ea hau e romelloa, u tla fumana lengolo-tsoibila le nang le nomoro ea ho batla le sehokelo sa ho latela thomello ea hau.
U ka latela odara ea hau ka kotloloho webosaeteng ea rona ka ho kena akhaonteng ea hau mme u shebe nalane ea hau ea odara.
Lithibelo tsa ho tsamaisa
Ka kopo tlatsa aterese ea seterateng ka botlalo, eseng lebokose la Pox kapa aterese ea sesole (APP). Ho seng joalo, re tla tlameha ho sebelisa ems bakeng sa ho tsamaisa (e ea lieha ho feta tse ling, ho nka 1 - ho nka likhoeli tse peli kapa nako e telele.
Mesebetsi ea Litloaelo le Tafole ea Lekhetho
Ka kopo elelloa hore mesebetsi efe kapa efe ea lekhetho, lekhetho kapa litefo tsa ho tlisa thepa tse hlahisitsoeng nakong ea thomello ke boikarabello ba moreki. Litefiso tsena li fapana ho latela naha e eang 'me li khethoa ke ba boholong sebakeng sa lehae.
Ka ho re reka webosaeteng ea rona, o lumela ho lefa mesebetsi efe kapa efe e sebetsang kapa lekhetho le amanang le odara ea hau. Ha re ikarabelle bakeng sa ho lieha ho bakoang ke tumello ea litloaelo.
Phokotso ea "PUSELETSO EA PUSO
Hang ha odara ea hau e fihlile sebakeng se khethiloeng sa Pionep Point kapa sebaka sa ho tsamaisa, ka kopo netefatsa pokello ea potlako. Haeba sephutheloana se sa nkuoe ka nako e behiloeng, re tla romella khopotso ka lengolo-tsoibila kapa sms. Leha ho le joalo, haeba sephutheloana se sa bokelloa nakong e boletsoeng, 'me tahlehelo e teng kapa tšenyo e etsahala ka lebaka leo, moreki o tla jara boikarabelo. Re u hopotse ka mosa hore u bokelle pakete ea hau hang-hang ho qoba litaba life tse ka bang teng.
Tlhokomeliso: Ha sehlahisoa sa rona se oela tlasa sehlopha se khethehileng, ho khutla le theko ha se amoheloe.
