Yintoni i-polyacrylamide gel electrophoresis?

Polyacrylamide Gel Electrophoresis

I-Gel electrophoresis bubuchule obusisiseko kwiilabhoratri kuzo zonke iinkalo zebhayoloji, evumela ukwahlulwa kwama-macromolecules afana ne-DNA, i-RNA kunye neeproteni.Imithombo eyahlukeneyo yokwahlula kunye neendlela zivumela ii-subsets zezi molekyuli ukuba zihlulwe ngokufanelekileyo ngokuxhaphaza iimpawu zabo zomzimba.Kwiiprotheni ngokukodwa, i-polyacrylamide gel electrophoresis (PAGE) ihlala iyindlela yokuzikhethela.

I-PAGE yindlela yokwahlula ii-macromolecules ezifana neeprotheni ezisekelwe ekuhambeni kwazo kwe-electrophoretic, oko kukuthi, ukukwazi kwabahlalutyi ukuya kwi-electrode yentlawulo echaseneyo.KwiPHEPHA, oku kumiselwa ngumrhumo, ubungakanani (ubunzima bemolekyuli) kunye nokumila kwemolekyuli.Abahlalutyi bahamba ngeepores ezenziwe kwijeli ye-polyacrylamide.Ngokungafaniyo ne-DNA kunye ne-RNA, iiprotheyini ziyahluka ngokweentlawulo ngokwee-amino acids ezidityanisiweyo, ezinokuphembelela indlela eziqhuba ngayo.Imitya ye-amino acid inokuthi yenze izakhiwo zesibini ezichaphazela ubungakanani bazo obubonakalayo kwaye ngenxa yoko zikwazi njani ukuhamba ngeepores.Ngoko ke ngamanye amaxesha kunokuba lunqweneleka ukuba denature iiproteni phambi electrophoresis ukuba linearize ukuba uqikelelo oluchanekileyo ngakumbi ubungakanani kufuneka.

IPHEPHA le-SDS

I-sodium-dodecyl sulfate i-polyacrylamide gel electrophoresis yindlela esetyenziselwa ukwahlula iimolekyuli zeprotheni zobuninzi be-5 ukuya kwi-250 kDa.Iiprotheni zihlulwe kuphela ngesiseko sobunzima bemolekyuli.I-sodium dodecyl sulfate, i-anionic surfactant, yongezwa ekulungiseleleni iigels ezifihla iintlawulo zangaphakathi zeesampulu zeprotheni kwaye zinike intlawulo efanayo kumlinganiselo wobunzima.Ngamagama alula, i-denatures iiprotheyini kwaye ibanika intlawulo embi.

IPHEPHA lomthonyama

I-PAGE yoMthonyama bubuchule obusebenzisa iigels ezingezona-denatured zokwahlula iiprotheni.Ngokungafaniyo nePHEPHA le-SDS, akukho arhente ye-denaturing yongezwa ekulungiseleleni iigels.Ngenxa yoko, ukuhlukana kweeprotheni kwenzeka ngesiseko sentlawulo kunye nobukhulu beeprotheni.Kobu buchule, ukuhambelana, ukugoqa kunye namatyathanga e-amino acid yeeprotheni zizinto ezixhomekeke kuzo ukwahlukana.Iiprotheyini azonakaliswa kule nkqubo, kwaye ziyakwazi ukubuyiswa emva kokugqitywa kokuhlukana.

Isebenza njani i-polyacrylamide gel electrophoresis (PAGE)?

Umgaqo osisiseko wePAGE kukwahlula abahlalutyi ngokubadlulisa kwiipores zejeli yepolyacrylamide usebenzisa umsinga wombane.Ukufezekisa oku, i-acrylamide–bisacrylamide mix is ​​polymerized (polyacrylamide) ngokongezwa kwe-ammonium persulfate (APS).I-reaction, eyenziwa yi-tetramethylethylenediamine (TEMED), yenza umnatha onjengesakhiwo kunye nama-pores apho abahlalutyi banokuhamba (Umfanekiso 2).Ipesenteji ephezulu ye-acrylamide epheleleyo efakwe kwijeli, incinci isayizi yepore, ngoko ke iiprotheni ezincinci ziya kukwazi ukudlula.Umyinge we-acrylamide ukuya kwi-bisacrylamide uya kuba nefuthe kubungakanani bepore kodwa oku kuhlala kugcinwa kungatshintshi.Ubungakanani bemibhobho emincinci bukwanciphisa isantya apho iiproteni ezincinci zikwazi ukuhamba ngejeli, ziphucula ukusonjululwa kwazo kwaye zibathintele ukuba babaleke baye kwi-buffer ngokukhawuleza xa kusetyenziswa okwangoku.

Izixhobo zePolyacrylamide Gel Electrophoresis

Iseli yeGel Electrophoresis (Itanki/Igumbi)
Itanki yejeli ye-polyacrylamide gel electrophoresis (PAGE) yahlukile kwitanki yejeli ye-agarose.Itanki yejeli yeagarose ithe tye, ngelixa itanki yePAGE ime nkqo.Ngeseli ethe nkqo ye-electrophoresis (itanki/igumbi), ijeli ecekethekileyo (ngokuqhelekileyo yi-1.0mm okanye i-1.5mm) igalelwa phakathi kweepleyiti zeglasi ezimbini kwaye inyuswe ukuze umzantsi wejeli untywiliselwe kwi-buffer kwigumbi elinye kwaye umphezulu untywiliselwe kwi-buffer. kwelinye igumbi.Xa kusetyenziswa umsinga, isixa esincinci sesithinteli sihamba ngejeli ukusuka kwigumbi eliphezulu ukuya kwigumbi elisezantsi.Ngeebamba ezomeleleyo zokuqinisekisa ukuba indibano ihlala kwindawo ethe tye, isixhobo sincedisa ukubaleka kwejeli ngokukhawuleza nokupholisa okukhokelela kwiibhanti ezahlukileyo.

I-Beijing Liuyi Biotechnology Co., Ltd. (i-Liuyi Biotechnology) yenza uluhlu lweeseli ze-polyacrylamide gel electrophoresis (itanki / amagumbi).Iimodeli ze-DYCZ-20C kunye ne-DYCZ-20G ziiseli ze-electrophoresis ezithe nkqo (itanki / amagumbi) kuhlalutyo lokulandelelana kwe-DNA.Ezinye zeeseli ze-electrophoresis ezithe nkqo (iitanki / amagumbi) zihambelana nenkqubo yokuqhawula, njengemodeli ye-DYCZ-24DN, i-DYCZ-25D kunye ne-DYCZ-25E iyahambelana ne-Western Blotting imodeli yenkqubo ye-DYCZ-40D, i-DYCZ-40G kunye ne-DYCZ-40F, ezisetyenziselwa ukuhambisa imolekyuli yeprotheni ukusuka kwijeli ukuya kwinwebu.Emva kwe-SDS-PAGE electrophoresis, i-Western Blotting yindlela yokufumanisa iprotheni ethile kumxube weprotheni.Unokukhetha ezi nkqubo zokuvala ngokweemfuno zovavanyo.

Electrophoresis Power Supply
Ukubonelela ngombane wokuqhuba ijeli, uya kufuna i-electrophoresis power supply.Kwi-Liuyi Biotechnology sibonelela ngoluhlu lwamandla ombane we-electrophoresis kuzo zonke izicelo.Imodeli ye-DYY-12 kunye ne-DYY-12C ene-voltage ephezulu ezinzileyo kunye nekhoyo ingahlangabezana nemfuneko yombane ophezulu we-electrophoresis.Inomsebenzi wokuma, ixesha, i-VH kunye nesinyathelo-ngesinyathelo isicelo.Zifanelekile kwi-IEF kunye ne-DNA yokulandelelanisa isicelo se-electrophoresis.Kwiprotheni jikelele kunye nesicelo se-electrophoresis ye-DNA, sinemodeli ye-DYY-2C, i-DYY-6C, i-DYY-10, njalo njalo, eziye zithengisa umbane oshushu kunye neeseli ze-electrophoresis (itanki / amagumbi).Ezi zinokusetyenziselwa usetyenziso lwe-electrophoresis yombane ophakathi kunye nophantsi, njengosetyenziso lwelebhu yesikolo, ilebhu yasesibhedlele njalo njalo.Iimodeli ezininzi zokubonelela ngombane, nceda undwendwele iwebhusayithi yethu.

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Iireferensi ze Yintoni i-polyacrylamide gel electrophoresis?
1. Karen Steward PhD Polyacrylamide gel electrophoresis, Indlela Esebenza ngayo, Ukwahluka koBuchule, kunye nokusetyenziswa kwayo