Nigute ushobora gukoresha kole yo gutandukana?
Iburengerazuba blot membrane kwimura
Mubikorwa byubu, poroteyine yimurwa ivuye muri gel ikajyana mubitwara bikomeye (membrane).
Guhitamo Membrane: ibikoresho bisanzwe bikoreshwa mugice cyo gucapa harimo NC membrane, DBM, DDT, nylon membrane, PVDF, nibindi. Duhitamo PVDF (fluoride polyvinylidene fluoride), ifite poroteyine nziza, imbaraga z'umubiri hamwe no guhuza imiti neza.Hano haribisobanuro bibiri: immobilon-p (0.45um) na immobilon PSQ (0.2um kuri MW <20kDa).
Uburyo bwumye
Nukuvuga, gel interlayer ikomatanya ishyirwa hagati yurupapuro rwungurura hamwe na transfert yoherejwe, kandi ikigezweho ikorwa binyuze muri buffer yamamajwe ku mpapuro zo kuyungurura kugirango igere ku ngaruka zo kwimura.Kuberako ibyubu bikora kuri firime ya kole, ibintu byoherejwe birakaze, ariko igihe cyo kwimura ni gito kandi imikorere ni myinshi.
(1) guhitamo ibihe byubushakashatsi
Ibiriho ni 1ma-2ma / cm2, kandi mubisanzwe dukoresha 100mA / membrane.Igihe cyo kwimura gishobora gutoranywa ukurikije ingano ya molekile ya poroteyine yagenewe hamwe na gel hamwe, kandi birashobora guhinduka uko bikwiye ukurikije uko ibintu bimeze.
Intego ya poroteyine ingana (kDa), kwibanda kwa gel hamwe nigihe cyo kohereza
80 --- 140 8% 1.5-2.0
25 --- 80 10% 1.5
15-40 12% 0,75
<20 15% 0.5
(2) imikorere yubushakashatsi
(1) Gutegura impapuro zungurura na membrane (imyiteguro igomba gutangira iminota 20 mbere ya electrophoreis).
A. Reba niba hari transfers zihagije, zitateguwe ako kanya.
B. Reba niba hari akayunguruzo impapuro na membrane yubunini bukwiye.
C. Shira membrane muri methanol muminota igera kuri 1-2.Hanyuma hanyuma yimurirwa muri transfert yoherejwe.
D. Shira impapuro zikwiye zifata impapuro hamwe na membrane iyungurura impapuro muri transfert yoherejwe.
PDVF membrane igomba gushiramo methanol kugeza ryari mugihe winjiye muri transfert?
PVDF ni hydrophobique, kandi biragoye gushira mumashanyarazi.Nyuma yo kuvura methanol, biroroshye gushiramo.Kwiyitirira PVDF nugushiramo no kuyikorana na methanol, hanyuma ukakaraba kabiri n'amazi yatoboye nyuma yo koga neza.Intego yo gukoresha methanol bubbles nugukora amatsinda yashizwemo neza kuri membrane ya PVDF no koroshya guhuza na proteyine zashizwemo nabi.
Abantu bakunze kuyikoresha muminota 5-20.Irashobora gushiramo icyarimwe no gufata kole, kandi bivugwa ko bizatwara iminota 5.Ingaruka ntabwo ari mbi.Mubihe byashize, inshuti zimwe zurubuga zanditse ko igihe cyo gutunganya ntacyo cyatwaye, ariko urufunguzo ni ubwiza bwa firime.Nibyo.Igihe cyose ururenda rwuzuye, bigomba kuba byiza.
Ibisobanuro bya PVDF ya Hybond isoma mu buryo bukurikira: "banza utose membrane muri 100% metanol (amasegonda 10)".Ndabyumva nuko Nibyiza gushiramo byibuze amasegonda 10.Mubyukuri, Nibyiza gushiramo amasegonda 10 cyangwa iminota 10.
(2) Kwimura
A. Shira impapuro zo hasi zo kuyungurura ku gikoresho cyohereza amashanyarazi.Mubisanzwe ibice bitatu bikoreshwa.
B. Shyira ururenda ku mpapuro zungurura urwanya ururondogoro, witondere kutagira ibibyimba hagati ya membrane nimpapuro zungurura, hanyuma usukemo buffer zoherejwe kuri membrane kugirango utume ururenda rutose.
C. Kuramo kole, kura geli ya stack hanyuma uyitondere witonze kuri firime.
D. Kata imfuruka yo hejuru yibumoso ya firime hanyuma ushire akamenyetso kuri kole kuri firime n'ikaramu.
E. Gupfundikanya urupapuro rwiyungurura urwanya kole.Suka bimwe byoherejwe hanyuma ushireho ibice bibiri byunguruzo.
F. Shyiramo ibikoresho byohereza amashanyarazi hanyuma uhitemo ibikenewe hamwe nigihe ukurikije ibikenewe.
G. Mugihe cyo kwimura, impinduka za voltage zigomba kugaragara igihe icyo aricyo cyose, kandi nibidasanzwe bizahindurwa mugihe.
Igihe cyo kohereza: Nzeri-07-2022
