Indlela yokusebenzisa iglue yokwahlula?–Icandelo 1

Indlela yokusebenzisa iglue yokwahlula?

Ukudluliselwa kwenwebu ye-blot yaseNtshona

Ngaphantsi kwesenzo sangoku, iprotheni idluliselwa kwi-gel kwi-carrier carrier (imembrane).

Ukukhetha i-Membrane: izinto eziqhelekileyo ezisetyenziswayo eziqinileyo-isigaba sokushicilela ziquka i-NC membrane, i-DBM, i-DDT, i-nylon membrane, i-PVDF, njl.Kukho iinkcukacha ezimbini: immobilon-p (0.45um) kunye ne-immobilon PSQ (0.2um ye-MW <20kDa).

Indlela eyomileyo

Oko kukuthi, indibaniselwano ye-gel interlayer ibekwe phakathi kwephepha lokucoca kunye ne-buffer yokudlulisa, kwaye yangoku iqhutywe nge-buffer adsorbed kwiphepha lokucoca ukufezekisa umphumo wokudlulisa.Ngenxa yokuba izenzo zangoku ngokuthe ngqo kwi-glue yefilimu, iimeko zokudlulisa zinobunzima, kodwa ixesha lokudlulisa lifutshane kwaye ukusebenza kakuhle kuphezulu.

(1) ukhetho lweemeko zovavanyo

Okwangoku yi-1ma-2ma / cm2, kwaye ngokuqhelekileyo sisebenzisa i-100mA / inwebu.Ixesha lokudlulisa linokukhethwa ngokobungakanani bemolekyuli yeprotheyini ekujoliswe kuyo kunye ne-concentration ye-gel, kwaye inokulungiswa ngokufanelekileyo ngokwemeko yangempela.

Iprotheyini ekujoliswe kuyo ubukhulu bemolekyuli (kDa), i-gel concentration kunye nexesha lokudlulisa

80---140 8% 1.5-2.0

25---80 10% 1.5

15—40 12% 0,75

<20 15% 0.5

(2) umsebenzi wokulinga

(1) Ukulungiswa kwephepha lokucoca kunye ne-membrane (ukulungiswa kuya kuqaliswa kwimizuzu engama-20 ngaphambi kwe-electrophoresis).

A. Jonga ukuba ngaba kukho izithinteli zodluliselo ezaneleyo, ezingalungiswanga ngoko nangoko.

B. Jonga ukuba kukho iphepha lokucoca kunye nenwebu yobukhulu obufanelekileyo.

C. Gxininisa inwebu kwi-methanol malunga nemizuzu eyi-1-2.Kwaye emva koko idluliselwe kwisithinteli sodluliselo.

D. Txisa iphepha lokucoca elincamathelayo elifanelekileyo kunye nephepha lokucoca inwebu kwisithinteli sokudlulisa ngokulandelelanayo.

Kufuneka inwebu yePDVF ihlale ixesha elingakanani kwimethanol xa ingena kwisithinteli sokudlulisa?

I-PVDF i-hydrophobic, kwaye kunzima ukufunxa kwisithinteli sokudlulisa.Emva konyango lwe-methanol, kulula ukucwina.Ukunyangwa kwangaphambili kwe-PVDF kukufunxa kwaye kusebenze nge-methanol, kwaye emva koko ihlambe kabini ngamanzi adibeneyo emva kokucwiliswa kakuhle.Injongo yokusebenzisa i-methanol bubbles kukuvula amaqela ahlawulwe kakuhle kwi-membrane ye-PVDF kwaye kube lula ukubopha ngeeprotheni ezihlawuliswa kakubi.

Abantu badla ngokuyisebenzisa imizuzu emi-5 ukuya kwengama-20.Inokuthi ifakwe ngexesha elifanayo nokuthatha iglue, kwaye kuqikelelwa ukuba kuya kuthatha malunga nemizuzu emi-5.Isiphumo asibi.Ngaphambili, abanye abahlobo bewebhu bathumele ukuba ixesha lokucubungula alinandaba kangako, kodwa eyona nto yayiyikhwalithi yefilimu.Yinyani leyo.Ngethuba nje i-membrane ifakwe ngokupheleleyo, kufuneka ilungile.

Inkcazo ye-PVDF ye-Hybond ifundeka ngolu hlobo: "ngaphambi kokumanzisa inwebu nge-100% ye-metanol (imizuzwana eyi-10)".Ukuqonda kwam kukuba kulungile ukucwina imizuzwana eyi-10.Enyanisweni, kulungile ukucwina imizuzwana eyi-10 okanye imizuzu eyi-10.

(2) Ukudluliselwa

A. Beka iphepha lokucoca elisezantsi kwisixhobo sokudlulisa umbane.Ngokuqhelekileyo kusetyenziswa iileya ezintathu.

B. Beka inwebu kwiphepha lokucoca ngokuchasene nenwebu, nikela ingqalelo ukuba ungabi namaqamza phakathi kwenwebu kunye nephepha lokucoca, kwaye ugalele isithinteli sokudlulisa kwinwebu ukugcina inwebu imanzi.

C. Gcoba i-glue, susa i-gel stack kwaye uyihambise ngokucophelela kwifilimu.

D. Sika ikona eliphezulu lasekhohlo lefilimu kwaye uphawule indawo yeglue kwifilimu ngepensile.

E. Gubungela isiqwenga sephepha lokucoca ngokuchasene neglu kwiglu.Galela isithinteli sokudlulisa kwaye ubeke amaqhekeza amabini ephepha lokucoca.

F. Faka isixhobo sokudlulisa umbane kwaye ukhethe okwangoku kunye nexesha elifunekayo ngokweemfuno.

G. Ngethuba lenkqubo yokudlulisa, utshintsho lombane luya kuqwalaselwa nangaliphi na ixesha, kwaye nayiphi na into engaqhelekanga iya kulungiswa ngexesha.