Kungani i-Polyacrylamide isetshenziselwa i-protein electrophoresis

Emkhakheni wesayensi yesimanje, i-protein electrophoresis iyindlela eyinhloko yokuhlaziya nokuchaza amaprotheni. Inhliziyo yale ndlela ileleI-Polyacrylamide, inhlanganisela eguquguqukayo esebenza njengomgogodla wama-matrices e-gel asetshenziswa ezinhlelweni ze-gel electrophoresis. Izakhiwo eziyingqayizivele ze-Polyacrylamide zenza kube ithuluzi elibalulekile kubacwaningi nososayensi abafuna ukuxazulula ubunzima bamaprotheni kanye nokusebenzisana kwawo.

I-Polyacrylamide, evame ukubizwa ngokuthi i-PAM, iyi-polymer yokwenziwa eyenziwe ngama-acrylamide monomers. Ukuguquguquka kwayo okuphawulekayo kubangelwa ikhono layo lokwenza amaketanga amade, okuholela entweni efana nejeli engamukela osayizi abahlukahlukene bama-molecule. Lesi sici senza i-polyacrylamide ibe yi-candidate efanelekile yokudala ama-matrices anezimbobo asetshenziswa ku-electrophoresis yamaprotheni.

I-protein electrophoresis iyindlela ehlukanisa amaprotheni ngokusekelwe ekushajeni kwawo kanye nosayizi wawo. Ngokufaka isampula yamaprotheni ensimini kagesi ngaphakathi kwe-polyacrylamide gel matrix, amaprotheni ahamba ngejeli ngamazinga ahlukene, okuholela kuma-band ahlukene angahlaziywa futhi alinganiswe. Lokhu kuhlukaniswa kunikeza ukuqonda okubalulekile ngobumsulwa bamaprotheni, ukunqunywa kwesisindo sama-molecule, kanye nokuba khona kwama-isoform.

Indima yePolyacrylamide ku-Protein Electrophoresis

Ukukhethwa kwe-polyacrylamide ye-protein electrophoresis kusekelwe emvelweni wayo oguquguqukayo. Ososayensi bangalungisa ukuhlushwa kwe-gel matrix ukuze kuhlangatshezwane namaprotheni anobukhulu obuhlukene. Ukuhlushwa okuphezulu kudala ama-matrices aqinile afanelekela ukuxazulula amaprotheni amancane, kuyilapho ukuhlushwa okuphansi kusetshenziswa amaprotheni amakhulu. Lokhu kuvumelana nezimo kuqinisekisa ukuthi abacwaningi bangakwazi ukwenza izivivinyo zabo zivumelane ukuze kufezwe ukuhlukaniswa nokuhlaziywa okuhle kakhulu.

I-Polyacrylamide njenge-I-Flocculant

Ukusetshenziswa kwe-Polyacrylamide kudlulela ngale kwendima yayo ku-gel electrophoresis. Iphinde ithole ukusetshenziswa njenge-flocculant ezimbonini ezahlukahlukene, njengokwelashwa kwamanzi kanye nokuphathwa kwamanzi amdaka. Njenge-flocculant, i-polyacrylamide iyasiza ekuhlanganiseni izinhlayiya ezilengayo koketshezi, okwenza kube lula ukususwa kwazo. Lesi sici sigqamisa amakhono ahlukahlukene ale nhlanganisela kanye nomthelela obanzi kwisayensi kanye nemboni.

Intuthuko ku-Electrophoresis Esekelwe ku-Polyacrylamide

Iminyaka yamuva nje ibone intuthuko eqhubekayo kumasu e-electrophoresis asekelwe ku-polyacrylamide. I-Native PAGE, i-SDS-PAGE, kanye ne-electrophoresis ye-gel enezinhlangothi ezimbili ziyizibonelo ezimbalwa nje zendlela ukuzivumelanisa nezimo kwe-polyacrylamide okuye kwenza ngayo ukuthuthukiswa kwezindlela ezikhethekile zokuhlaziya izakhiwo zamaprotheni, ukuguqulwa kwangemva kokuhumusha, kanye nokusebenzisana. Lezi zindlela zibaluleke kakhulu ocwaningweni lwe-proteomics kanye nemizamo yokuthola izidakamizwa.

Emkhakheni wokuhlaziywa kwamaprotheni, i-polyacrylamide ivela njengomngane oqinile, okwenza abacwaningi bakwazi ukungena ezweni eliyinkimbinkimbi lamaprotheni. Indima yayo njengesisekelo se-gel matrices ezinhlelweni ze-electrophoresis ayinakweqiswa. Kusukela ekuhlukaniseni izindlela zezifo kuya ekuthuthukiseni izindlela zokwelapha ezintsha, i-electrophoresis esekelwe ku-polyacrylamide iyaqhubeka nokubumba intuthuko yesayensi. Njengoba ubuchwepheshe buthuthuka, lesi simangaliso sokwenziwa cishe sizothuthuka, sithuthukise ukuqonda kwethu amaprotheni nemisebenzi yawo eminingi.