Kuyesa kulekanitsa mapuloteni a seramu ndi Cellulose Acetate Membrane Electrophoresis

Mfundo yofunika

Cellulose acetate film electrophoresis ndi njira ya electrophoresis pogwiritsa ntchito filimu ya cellulose acetate monga chithandizo. Chodabwitsa chomwe tinthu tating'onoting'ono timapita ku mbali ina ya elekitirodi pansi pa mphamvu ya magetsi amatchedwa electrophoresis. Popeza puloteni iliyonse imakhala ndi malo enieni a isoelectric, ngati puloteni imayikidwa mu njira yothetsera pH yotsika kusiyana ndi isoelectric point, puloteniyo imayendetsedwa bwino ndikusunthira ku electrode yolakwika. M'malo mwake, imasunthira kumtengo wabwino. Chifukwa liwiro la mamolekyu a protein omwe akuyenda m'munda wamagetsi amagwirizana ndi mtengo wawo, mawonekedwe ndi kukula kwa mamolekyu, mapuloteni osiyanasiyana amatha kulekanitsidwa ndi electrophoresis. Seramu imakhala ndi mapuloteni osiyanasiyana, onse omwe ali ndi isoelectric point pa pH7.5 kapena kuchepera. Seramu ikayikidwa mu pH 8.6 buffer kuti igwiritse ntchito electrophoresis, mapuloteni onse a seramu amakhala olakwika ndipo amasunthira ku mbali yabwino yamagetsi. Popeza mapuloteni osiyanasiyana a seramu amakhala ndi milandu yosiyana pa pH yomweyo, tinthu tating'onoting'ono timasiyana kukula kwake, motero liwiro la kusamuka ndi losiyana, ndipo amasiyanitsidwa ndi electrophoresis. Ma isoelectric point ndi kulemera kwa mamolekyu a mapuloteni a seramu akuwonetsedwa patebulo ili pansipa:

Kuyesera ndikulekanitsa mapuloteni osiyanasiyana mu seramu yamagazi yokhala ndi cellulose acetate membrane (abbr. CAM) ngati media yothandizira. CAM ndi mtundu wa loos wa thovuefilimu yokhala ndi yunifolomu yabwino, ndipo makulidwe ake ndi 0.1mm-1.5mm, omwe ali ndi mayamwidwe ena amadzi.

Zida, zida ndi ma reagents a CAM Electrophoresis

Zitsanzo:seramu yamagazi amunthu wathanzi

Chida:magetsi DYY-6C, thanki electrophoresis DYCP-38C, wapamwamba chitsanzo potsegula WD-9404

The reagents

1) cellulose acetate nembanemba 7X9cm

2) PH 8.6 barbitol buffer solution (ionic mphamvu 0.05-0.09, ndi nthawi ya 0.06tid.): tengani 1,84g Diethyl barbituric acid, kenako tengani 1.03g Diethyl sodium pentobarbital, onjezerani madzi osungunuka kuti mutenthe kuti asungunuke, ndikupanga mpaka 1000 ml;

3) Kuthimbirira: Ponceau S 0.2g, Trichloroacetic 3g, kuwonjezera 100ml madzi osungunuka;

4) TBS / T kapena PBS / T: 45ml 95% ethanol, 5ml glacial acetic acid, kuwonjezera 50ml madzi osungunuka;

5) Kuyeretsa Njira: 70ml anhydrous ethanol, 30ml glacial acetic acid.

Yesani Njirad

1) Konzani nembanemba: Mizirani nembanemba mu njira ya barbitol buffer 30min-8h, ndikuitulutsa, chotsani njira yowonjezerapo ndi pepala loyamwa.

2) Kuyika zitsanzo: Siyanitsani mbali yolimba ndi yosalala ya nembanemba, ndipo lembani mzere pa mtunda wa 1.5cm mpaka kumapeto kwa pamwamba pa mbali yovuta. Kwezani zitsanzo ndi chida chapamwamba chotsitsa chachitsanzo pambali yoyipa. Zindikirani: Zitsanzozi ziyenera kuyikidwa pambali yolimba ya membrane. Zitsanzo za seramu zitalowetsedwa mu nembanemba, tembenuzirani nembanemba, ikani mbali yoyipa (yokhala ndi zitsanzo) moyang'anizana ndi thanki, ndipo kumapeto kwake ndi zitsanzo kumayikidwa mu electrode yoyipa.

3) Electrophoresis: Yatsani magetsi, 0.4～0.6m A / masentimita membrane, nthawi yothamanga ndi 30-45 min. Pambuyo kuthamanga electrophoresis, zimitsani mphamvu.

4) Tsitsani ndi kuyeretsa: Chotsani nembanemba mu thanki, ndikumizait mu njira ya utoto kwa 5 min, ndiyeno muyeretseni mu njira yoyeretsera mobwerezabwereza kwa 3-4 mpaka mtundu wakumbuyo utawonekera. Mapuloteni a seramu ayenera kuwonetsedwa pamagulu, ndipo kawirikawiri pali madera asanu, kupanga mapeto apamwamba ku mzere wodziwika, Albumin, α1-gloulin, α2-gloulin, β-gloulin, γ-gloulin.

5) Kusungirako: ikani electrophoresis youmagulupoyeretsa yankho kwa mphindi 10-15, kenako ndikuitulutsa ndikuyiyika pagalasi loyera, ikauma, idzakhala filimu yowonekera.gulu.

YesaniZotsatira

Kusiyanitsa kwa zitsanzo za seramu ndikwabwino, ndipo palibe chodabwitsa cha band. Kubwereza kwa zotsatira kumasiyanasiyana chifukwa cha njira zoyesera ndi njira zoyesera, ndipo kubwereza ndikokwera.

Mapeto

Njira yodziwira matenda a electrophoresis mwachangu (Electrophoresis thankiDYCP-38C,magetsiDYY-6C ndi chitsanzo chapamwamba chotsitsa WD-9404) chopangidwa ndi athuKampani Beijing Liuyi Biotechnology Co., Ltdamakwaniritsa zofunikira zoyeserera,ndizotsatira zake ndi reproducible, zosavuta ndi mofulumira, oyenerakuphunzitsakafukufuku.

Beijing LiuyiBiotechnology Co., Ltd imakhazikika pakupanga zinthu za elecrtophoresis zamakampani a sayansi ya moyo, zomweili ndi mbiri yopitilira zaka 50 ku China ndipo kampaniyo imatha kupereka zinthu zokhazikika komanso zapamwamba padziko lonse lapansi. Kupyolera muzaka zachitukuko, ndizoyenera kusankha kwanu!

Tsopano tikuyang'ana mabwenzi, onse a OEM electrophoresis thanki ndi ogulitsa amalandiridwa.

Kuti mudziwe zambiri za ife, chonde titumizireni imelo[imelo yotetezedwa]kapena[imelo yotetezedwa].