Polyacrylamide gel electrophorei ni iki?

Polyacrylamide Gel Electrophoresis

Gel electrophorei ni tekinike yibanze muri laboratoire hirya no hino mubinyabuzima, yemerera gutandukanya macromolecules nka ADN, RNA na proteyine. Itangazamakuru ritandukanye hamwe nuburyo butandukanye butuma ibice bya molekile bitandukana neza mugukoresha ibiranga umubiri. Kuri poroteyine byumwihariko, polyacrylamide gel electrophorei (PAGE) akenshi ni tekinike yo guhitamo.

URUPAPURO ni tekinike itandukanya macromolecules nka poroteyine zishingiye ku kugenda kwa electrophoreque, ni ukuvuga ubushobozi bwa analyite bwo kwerekeza kuri electrode yumuriro utandukanye. URUPAPURO, ibi bigenwa nubushakashatsi, ingano (uburemere bwa molekile) nuburyo bwa molekile. Isesengura rinyura mu byobo byakozwe muri gel polyacrylamide. Bitandukanye na ADN na RNA, poroteyine ziratandukanye bitewe na aside amine yashizwemo, ishobora guhindura uko ikora. Imigozi ya acide ya Amino irashobora kandi gukora urwego rwa kabiri rugira ingaruka kubunini bwarwo bityo nuburyo bashoboye kunyura mumyenge. Birashobora rero rimwe na rimwe kwifuzwa gutandukanya poroteyine mbere ya electrophoreis kugirango zihuze umurongo niba hakenewe igereranya ryukuri ry'ubunini.

URUPAPURO RWA SDS

Sodium-dodecyl sulfate polyacrylamide gel electrophorei ni tekinike ikoreshwa mu gutandukanya molekile za poroteyine za misa 5 kugeza 250 kDa. Poroteyine zitandukanijwe gusa zishingiye ku buremere bwazo. Sodium dodecyl sulfate, anionic surfactant, yongewemo mugutegura geles zipfundikanya amafaranga yimbere yintungamubiri za poroteyine kandi ikabaha amafaranga asa numubare rusange. Mu magambo yoroshye, yerekana poroteyine kandi ikabaha amafaranga mabi.

URUPAPURO Kavukire

PAGE kavukire ni tekinike ikoresha geles idatandukanijwe kugirango itandukane na poroteyine. Bitandukanye na SDS PAGE, ntamukozi wongeyeho wongeyeho mugutegura geles. Nkigisubizo, gutandukanya poroteyine bibaho hashingiwe ku kwishyurwa no ku bunini bwa poroteyine. Muri ubu buhanga, guhinduka, kuzinga hamwe na aminide acide ya amine ya poroteyine nibyo bintu gutandukana biterwa. Poroteyine ntabwo zangiritse muriki gikorwa, kandi zirashobora kugarurwa nyuma yo gutandukana.

Nigute polyacrylamide gel electrophorei (PAGE) ikora?

Ihame ryibanze rya PAGE nugutandukanya analyite ubinyujije mumyobo ya gel polyacrylamide ukoresheje amashanyarazi. Kugirango ubigereho, kuvanga acrylamide - bisacrylamide ni polymerized (polyacrylamide) hiyongereyeho ammonium persulfate (APS). Imyitwarire, iterwa na tetramethylethylenediamine (TEMED), ikora urushundura nkimiterere ifite imyenge inyuramo analyite ishobora kunyuramo (Ishusho 2). Kurenza ijanisha rya acrylamide yose yashyizwe muri gel, niko ingano ya pore, bityo proteine ​​ntoya izashobora kunyuramo. Ikigereranyo cya acrylamide na bisacrylamide nacyo kizagira ingaruka ku bunini bwa pore ariko ibi bikomeza guhoraho. Ingano ntoya nayo igabanya umuvuduko poroteyine nto zishobora kunyura muri gel, zinonosora imikemurire yazo kandi zikababuza gusohoka muri buffer byihuse mugihe ikoreshwa.

Ibikoresho bya Polyacrylamide Gel Electrophoresis

Akagari ka Gel Electrophoresis (Tank / Urugereko)
Ikigega cya gel ya polyacrylamide gel electrophorei (PAGE) itandukanye na agarose gel. Ikigega cya agarose gel gitambitse, mugihe ikigega cya PAGE gihagaritse. Na vertical electrophoresis selile (tank / chamber), gel yoroheje (mubisanzwe 1.0mm cyangwa 1.5mm) isukwa hagati yamasahani abiri yikirahure hanyuma igashyirwaho kuburyo hepfo ya jel yarohamye muri buffer mucyumba kimwe hanyuma hejuru ikarohama muri buffer mu kindi cyumba. Iyo ikigezweho gishyizwe mubikorwa, umubare muto wa buffer wimuka unyuze muri gel uva mucyumba cyo hejuru ugana mucyumba cyo hasi. Hamwe na clamps ikomeye kugirango yizere ko inteko iguma mumwanya uhagaze, ibikoresho byorohereza gel byihuse hamwe no gukonjesha bivamo imirongo itandukanye.

Pekin Liuyi Biotechnology Co., Ltd. Moderi DYCZ-20C na DYCZ-20G ni selile vertical electrophoreis selile (tanks / chambers) kugirango isesengura ADN ikurikirana. Zimwe mu ngirabuzimafatizo za vertical electrophoreis (tanks / chambers) zirahujwe na sisitemu yo guhanagura, nka moderi DYCZ-24DN, DYCZ-25D na DYCZ-25E ihujwe na moderi ya Western Blotting ya DYCZ-40D, DYCZ-40G na DYCZ-40F, zikoreshwa mu kwimura molekile ya poroteyine kuva muri gel ikajya muri membrane. Nyuma ya SDS-PAGE electrophoreis, Western Blotting nubuhanga bwo kumenya poroteyine yihariye ivanze na poroteyine. Urashobora guhitamo sisitemu yo guhanagura ukurikije ibisabwa byubushakashatsi.

Amashanyarazi
Kugirango utange amashanyarazi yo gukoresha gel, uzakenera amashanyarazi ya electrophoreis. Kuri Liuyi Biotechnology dutanga urutonde rwamashanyarazi ya electrophoreis kubisabwa byose. Icyitegererezo DYY-12 na DYY-12C hamwe na voltage ihamye kandi ihindagurika irashobora kuzuza amashanyarazi menshi asabwa na electrophorei. Ifite imikorere yo guhagarara, igihe, VH hamwe nintambwe ku ntambwe. Nibyiza kuri IEF na ADN ikurikirana ya electrophoreis ikoreshwa. Kuri poroteyine rusange hamwe na ADN ya electrophoreis ikoreshwa, dufite icyitegererezo DYY-2C, DYY-6C, DYY-10, nibindi, nabyo bikaba ari ibikoresho byo kugurisha bishyushye hamwe na selile ya electrophoreis (tanks / chambers). Ibi birashobora gukoreshwa hagati ya voltage na electrophoresis ya voltage yo hagati, nko gukoresha laboratoire yishuri, laboratoire yibitaro nibindi. Moderi nyinshi zo gutanga amashanyarazi, nyamuneka sura urubuga rwacu.

Ikirango cya Liuyi gifite amateka yimyaka irenga 50 mubushinwa kandi isosiyete irashobora gutanga ibicuruzwa bihamye kandi byujuje ubuziranenge kwisi yose. Binyuze mumyaka yiterambere, birakwiye guhitamo!

Kubindi bisobanuro kuri twe, nyamuneka twandikire ukoresheje imeri[imeri irinzwe] or [imeri irinzwe].

Reba kuri polyacrylamide gel electrophorei ni iki?
1. Karen Igisonga cya PhD Polyacrylamide gel electrophorei, Uburyo ikora, Impinduka za tekinike, hamwe nibisabwa.