Iyini i-polyacrylamide gel electrophoresis?

I-Polyacrylamide Gel Electrophoresis

I-Gel electrophoresis iyindlela eyisisekelo kulabhorethri kuyo yonke imikhakha yebhayoloji, evumela ukuhlukaniswa kwama-macromolecules njenge-DNA, i-RNA namaprotheni. Imidiya ehlukene yokuhlukanisa nezindlela zivumela amasethi angaphansi kwalawa ma-molecule ukuthi ahlukaniswe ngokuphumelelayo ngokusebenzisa izici zawo zomzimba. Kumaprotheni ikakhulukazi, i-polyacrylamide gel electrophoresis (PAGE) imvamisa iyisu elikhethwayo.

I-PAGE yindlela ehlukanisa ama-macromolecules afana namaprotheni ngokusekelwe ekuhambeni kwawo kwe-electrophoretic, okungukuthi, ikhono labahlaziyi ukuya ku-electrode yenkokhiso ephambene. Ku-PAGE, lokhu kunqunywa ukushajwa, usayizi (isisindo se-molecule) kanye nokuma kwe-molecule. Abahlaziyi bahamba ngezimbotshana ezakhiwe kujeli ye-polyacrylamide. Ngokungafani ne-DNA ne-RNA, amaprotheni ayahlukahluka ngokuya ngama-amino acid ahlanganisiwe, angathonya indlela asebenza ngayo. Izintambo ze-amino acid zingase futhi zakhe izakhiwo zesibili ezithinta usayizi wazo osobala futhi ngenxa yalokho zikwazi ukuhamba kanjani ezimbotsheni. Ngakho-ke ngezinye izikhathi kungase kube okufiselekayo ukukhipha amaprotheni ngaphambi kwe-electrophoresis ukuze ahleleke uma kudingeka isilinganiso esinembe kakhudlwana sosayizi.

IKHASI le-SDS

I-sodium-dodecyl sulfate polyacrylamide gel electrophoresis iyindlela esetshenziselwa ukuhlukanisa ama-molecule wamaprotheni obuningi 5 kuya ku-250 kDa. Amaprotheni ahlukaniswa kuphela ngesisekelo sesisindo samangqamuzana awo. I-sodium dodecyl sulfate, i-anionic surfactant, yengezwa ekulungiseni amajeli avala izinkokhiso zangaphakathi zamasampula amaprotheni futhi iwanikeze inkokhiso efanayo nesilinganiso sobuningi. Ngamazwi alula, i-denatures amaprotheni futhi iwanikeze icala elibi.

I-PAGE yomdabu

I-Native PAGE yindlela esebenzisa amajeli angewona awe-denatured ekuhlukaniseni amaprotheni. Ngokungafani ne-SDS PAGE, ayikho i-ejenti ye-denaturing eyengezwa ekulungiseni ama-gel. Ngenxa yalokho, ukuhlukaniswa kwamaprotheni kwenzeka ngesisekelo sokukhokhiswa nobukhulu bamaprotheni. Kule nqubo, ukuhlanganisa, ukugoqa kanye namaketanga e-amino acid amaprotheni yizinto ukuhlukana kuncike kuzo. Amaprotheni awonakele kule nqubo, futhi angabuyiselwa ngemva kokuqedwa kokuhlukaniswa.

Isebenza kanjani i-polyacrylamide gel electrophoresis (PAGE)?

Umgomo oyisisekelo we-PAGE ukuhlukanisa ama-analyte ngokuwadlulisela ezimbotsheni zejeli ye-polyacrylamide kusetshenziswa amandla kagesi. Ukufeza lokhu, ingxube ye-acrylamide–bisacrylamide yenziwa i-polymerized (i-polyacrylamide) ngokufakwa kwe-ammonium persulfate (APS). Ukusabela, okwenziwe yi-tetramethylethylenediamine (TEMED), kwakha inetha elifana nesakhiwo esinezimbotshana lapho ama-analyte anganyakaza (Umfanekiso 2). Ukuphakama kwephesenti lengqikithi ye-acrylamide efakwe kujeli, kuba yincane usayizi wembotshana, yingakho kuba mancane amaprotheni azokwazi ukudlula. Isilinganiso se-acrylamide ne-bisacrylamide sizophinde sibe nomthelela kusayizi wezimbotshana kodwa lokhu kuvame ukugcinwa njalo. Izimbotshana ezincane ezimbotshana ziphinde zehlise isivinini lapho amaprotheni amancane akwazi khona ukuhamba ejeli, athuthukise ukucaca kwawo futhi awavimbele ukuthi abaleke aye ku-buffer ngokushesha lapho kusetshenziswa amandla.

Izisetshenziswa ze-Polyacrylamide Gel Electrophoresis

Iseli ye-Gel Electrophoresis (Ithangi/Igumbi)
Ithangi lejeli le-polyacrylamide gel electrophoresis (PAGE) lihlukile kuthangi lejeli ye-agarose. Ithangi lejeli le-agarose livundlile, kuyilapho ithangi le-PAGE limile. Ngeseli eqondile ye-electrophoresis (ithangi/igumbi), ijeli elincanyana (imvamisa engu-1.0mm noma u-1.5mm) lithelwa phakathi kwamapuleti amabili engilazi futhi lifakwe ukuze kucwiliswe ngaphansi kwejeli ku-buffer ekamelweni elilodwa futhi phezulu kucwiliswe ku-buffer. kwelinye ikamelo. Uma kusetshenziswa i-current, inani elincane le-buffer lihamba ngejeli lisuka ekamelweni eliphezulu liye ekamelweni elingaphansi. Ngama-clamp aqinile okuqinisekisa ukuthi ukuhlangana kuhlala kumile, okokusebenza kusiza ukugijima kwejeli ngokupholisa okulinganayo okuholela kumabhendi ahlukile.

I-Beijing Liuyi Biotechnology Co., Ltd. (i-Liuyi Biotechnology) ikhiqiza osayizi abahlukahlukene be-polyacrylamide gel electrophoresis cell (mathange/amagumbi). Izinhlobo ze-DYCZ-20C ne-DYCZ-20G zingamaseli e-electrophoresis aqondile (amathangi/amakamelo) okuhlaziya ukulandelana kwe-DNA. Amanye amaseli e-electrophoresis aqondile (amathangi/amagumbi) ahambisana nohlelo lokucisha, njengemodeli i-DYCZ-24DN, i-DYCZ-25D ne-DYCZ-25E iyahambisana ne-Western Blotting system model DYCZ-40D, DYCZ-40G kanye ne-DYCZ-40F, ezisetshenziselwa ukudlulisa i-molecule yamaprotheni isuka kujeli iye kulwelwesi. Ngemuva kwe-SDS-PAGE electrophoresis, i-Western Blotting iyindlela yokuthola iphrotheni ethile kwingxube yephrotheni. Ungakhetha lawa masistimu wokusula ngokuya ngezidingo zokuhlola.

I-Electrophoresis Power Supply
Ukuze unikeze ugesi wokusebenzisa ijeli, uzodinga amandla kagesi we-electrophoresis. Kwa-Liuyi Biotechnology sihlinzeka ngohlu lwamandla kagesi we-electrophoresis kuzo zonke izinhlelo zokusebenza. Imodeli ye-DYY-12 ne-DYY-12C enevoltheji ephezulu ezinzile futhi yamanje ingahlangabezana nemfuneko ye-electrophoresis ephezulu ye-voltage. Inomsebenzi wokuma, isikhathi, i-VH kanye nesicelo sesinyathelo ngesinyathelo. Zilungele i-IEF kanye ne-DNA yokulandelanisa i-electrophoresis application. Ngohlelo lokusebenza lwe-protein evamile ne-DNA electrophoresis, sinemodeli ye-DYY-2C, i-DYY-6C, i-DYY-10, njalonjalo, ephinde ibe amandla okuthengisa ashisayo anamaseli e-electrophoresis (mathangi/amagumbi). Lezi zingasetshenziselwa izinhlelo zokusebenza ze-electrophoresis ephakathi nephansi, njengokusebenzisa elebhu yesikole, ilebhu yasesibhedlela nokunye. Amamodeli engeziwe okunikezwa kwamandla, sicela uvakashele iwebhusayithi yethu.

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Izinkomba zokuthi Iyini i-polyacrylamide gel electrophoresis?
1. Ijeli lika-Karen Steward PhD Polyacrylamide gel electrophoresis, Indlela Esebenza Ngayo, Izinhlobonhlobo Zobuchule, kanye Nezinhlelo Zakho zokusebenza