Polyacrylamide gel Electrophoresis
Gel electrophoresis bụ usoro dị mkpa na ụlọ nyocha n'ofe usoro ihe ọmụmụ, na-enye ohere nkewa nke macromolecules dị ka DNA, RNA na protein. Usoro mgbasa ozi dị iche iche na usoro nkewa na-enye ohere ikewapụ akụkụ nke mkpụrụ ndụ ndị a nke ọma site n'iji njirimara anụ ahụ ha eme ihe. Maka protein karịsịa, polyacrylamide gel electrophoresis (PAGE) na-abụkarị usoro nhọrọ.
PAGE bụ usoro na-ekewa macromolecules dị ka protein na-adabere na ngagharị electrophoretic ha, ya bụ, ikike nke ndị nyocha ịkwaga n'ebe electrode nke ụgwọ dị iche. Na PAGE, a na-ekpebi nke a site na ụgwọ, nha (arọ molekụla) na ọdịdị nke molekul. Ndị nyocha na-aga site na pores etolite na gel polyacrylamide. N'adịghị ka DNA na RNA, protein na-adịgasị iche iche dị ka amino acid agbakwunyere, nke nwere ike imetụta otú ha si agba ọsọ. Eriri Amino acid nwekwara ike mebe ihe owuwu nke abụọ na-emetụta nha ha pụtara na ya mere otu ha si enwe ike isi na pores. Ya mere, ọ nwere ike na-achọsi ike mgbe ụfọdụ ka denature protein tupu electrophoresis linearize ha ma ọ bụrụ na a ọzọ ezi atụmatụ nke size chọrọ.
SDS PAGE
Sodium-dodecyl sulfate polyacrylamide gel electrophoresis bụ usoro eji kewaa protein ụmụ irighiri ihe dị ka 5 ruo 250 kDa. A na-ekewa protein ndị ahụ naanị na-adabere n'ịdị arọ molekụla ha. A na-agbakwunye sodium dodecyl sulfate, anonic surfactant, na nkwadebe nke gels nke na-ekpuchi ụgwọ dị n'ime nke ihe nlele protein ma na-enye ha ụgwọ yiri ya na oke oke. N'okwu dị mfe, ọ na-ewepụ protein ndị ahụ ma nye ha ụgwọ na-adịghị mma.
Ala ala PAGE
Native PAGE bụ usoro na-eji gels na-abụghị denatured maka nkewa nke protein. N'adịghị ka SDS PAGE, ọ dịghị onye na-edebanye aha na-agbakwunye na nkwadebe nke gels. N'ihi ya, nkewa nke protein na-ewere ọnọdụ na ndabere nke ụgwọ na size nke protein. Na usoro a, nchikota, mpịakọta na amino acid agbụ nke protein bụ ihe nkewa ahụ dabere na ya. Ndị na-edozi ahụ adịghị emebi emebi na usoro a, enwere ike nwetaghachi ya mgbe agwụchara nkewa.
Kedu ka polyacrylamide gel electrophoresis (PAGE) si arụ ọrụ?
Ụkpụrụ bụ isi nke PAGE bụ ikewapụ ndị nyocha site n'ịgafe ha site na pores nke gel polyacrylamide site na iji ọkụ eletrik. Iji mezuo nke a, a na-emepụta ngwakọta acrylamide-bisacrylamide (polyacrylamide) site na mgbakwunye nke ammonium persulfate (APS). Mmeghachi omume ahụ, nke tetramethylethylenediamine (TEMED) na-emezigharị, na-emepụta ụgbụ dị ka ihe owuwu nwere oghere nke ndị nyocha nwere ike isi na-ebugharị (Nyocha 2). Ka ọnụ ọgụgụ dị elu nke ngụkọta acrylamide gụnyere na gel, na-adị ntakịrị ntakịrị nha, ya mere obere protein ndị ga-enwe ike ịgafe. Oke nke acrylamide na bisacrylamide ga-emetụta oke pore mana nke a na-edobekarị. Obere pore nha na-ebelatakwa ọsọ nke obere ndị na-edozi ahụ na-enwe ike ịgafe na gel, na-eme ka mkpebi ha dịkwuo mma ma na-egbochi ha ịgbaba n'ime ihe nchekwa ngwa ngwa mgbe etinyere ugbu a.
Ngwa maka Polyacrylamide Gel Electrophoresis
Gel Electrophoresis Cell (Tank/Ụlọ)
Gel tank maka polyacrylamide gel electrophoresis (PAGE) dị iche na tank gel agarose. Agwarose gel tank bụ kehoraizin, ebe tank PAGE kwụ ọtọ. Site na cell electrophoresis vetikal (tank / ụlọ), a na-awụsa gel dị mkpa (nke na-adịkarị 1.0mm ma ọ bụ 1.5mm) n'etiti efere iko abụọ ma tinye ya ka ala nke gel wee banye n'ime ihe nchekwa n'otu ọnụ ụlọ ma n'elu na-emikpu n'ime ihe nchekwa. n'ọnụ ụlọ ọzọ. Mgbe etinyere ihe ugbu a, obere ihe nchekwa na-akwaga site na gel si n'ọnụ ụlọ dị n'elu gaa n'ọnụ ụlọ dị ala. Site na njigide siri ike iji kwe nkwa na mgbakọ ahụ ga-anọ n'ọnọdụ kwụ ọtọ, akụrụngwa na-eme ka gel na-agba ọsọ ọsọ na-eme ka ọ dị jụụ na-ebute eriri dị iche iche.
Beijing Liuyi Biotechnology Co., Ltd. (Liuyi Biotechnology) na-arụpụta ụdị dị iche iche nke polyacrylamide gel electrophoresis cell (tankị / ụlọ). Ụdị DYCZ-20C na DYCZ-20G bụ sel electrophoresis kwụ ọtọ (tankị / ụlọ) maka nyocha usoro DNA. Ụfọdụ n'ime vetikal electrophoresis cell (tankị / ụlọ) dakọtara na blotting usoro, dị ka ihe nlereanya DYCZ-24DN, DYCZ-25D na DYCZ-25E bụ dakọtara na Western Blotting usoro nlereanya DYCZ-40D, DYCZ-40G na DYCZ-40F. nke a na-eji maka ibufe molekul protein site na gel na akpụkpọ ahụ. Mgbe SDS-PAGE electrophoresis gasịrị, Western Blotting bụ usoro iji chọpụta otu protein dị na ngwakọta protein. Ị nwere ike ịhọrọ usoro nkwụsịtụ ndị a dịka ihe nyocha chọrọ.
Electrophoresis ike ọkọnọ
Iji nye ọkụ eletrik maka ịgbasa gel, ị ga-achọ ọkụ eletrik electrophoresis. Na Liuyi Biotechnology anyị na-enye ọtụtụ ọkụ eletrik maka ngwa niile. The nlereanya DYY-12 na DYY-12C na elu kwụsiri ike voltaji na ugbu a nwere ike izute elu voltaji chọrọ electrophoresis. Ọ nwere ọrụ nke nguzo, oge, VH na ngwa nzọụkwụ site na nzọụkwụ. Ha dị mma maka ngwa electrophoresis nke IEF na DNA. N'ihi na n'ozuzu protein na DNA electrophoresis ngwa, anyị nwere ihe nlereanya DYY-2C, DYY-6C, DYY-10, na na, bụ ndị na-ekpo ọkụ ahịa ike onunu na electrophoresis cell (tankị / ụlọ). Enwere ike iji ndị a maka ngwa electrophoresis nke etiti na nke dị ala, dị ka maka ụlọ nyocha ụlọ akwụkwọ, ụlọ ọgwụ ụlọ ọgwụ na ihe ndị ọzọ. Ụdị ndị ọzọ maka inye ọkụ, biko gaa na webụsaịtị anyị.
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Ntụaka maka Gịnị bụ polyacrylamide gel electrophoresis?
1. Karen Steward PhD Polyacrylamide gel electrophoresis, Olee otú o si arụ ọrụ, Usoro dị iche iche, na ngwa ya.
Oge nzipu: Mee-23-2022