Sabelane ngezinto ezimbalwa ezicatshangelwayo ngeviki eledlule zokusebenzisa i-cellulose acetate membrane electrophoresis, futhi sizoqedela lesi sihloko lapha namuhla ukuze uthole ireferensi yakho.
Ukukhethwa kwe I-Buffer Concentration
I-buffer concentration esetshenziswa ku-cellulose acetate membrane electrophoresis ngokuvamile iphansi kunaleyo esetshenziswa ku-electrophoresis yamaphepha. I-pH evame ukusetshenziswa ngu-8.6Bi-arbital buffer ngokuvamile ikhethwa phakathi kwebanga elingu-0.05 mol/L kuya ku-0.09 mol/L. Lapho kukhethwa ukuhlushwa, ukuzimisela kokuqala kwenziwa. Isibonelo, uma ubude be-membrane strip phakathi kwama-electrode ekamelweni le-electrophoresis buyi-8-10cm, i-voltage engu-25V iyadingeka ngesentimitha yobude be-membrane, futhi ukuqina kwamanje kufanele kube ngu-0.4-0.5 mA ngesentimitha yobubanzi be-membrane. Uma lawa manani engafinyelelwanga noma eqiwe ngesikhathi se-electrophoresis, ukugxila kwe-buffer kufanele kunyuswe noma kuhlanjululwe.
Ukugxiliswa kwebhafa okuphansi kakhulu kuzoholela ekuhambeni okusheshayo kwamabhendi kanye nokwanda kobubanzi bebhendi. Ngakolunye uhlangothi, ukugxiliswa kwebhafa ephezulu kakhulu kuzonciphisa ukufuduka kwebhendi, kwenze kube nzima ukuhlukanisa amabhendi athile okuhlukanisa.
Kufanele kuqashelwe ukuthi ku-cellulose acetate membrane electrophoresis, ingxenye ebalulekile yamanje iqhutshwa ngesampula, ekhiqiza inani elikhulu lokushisa. Kwesinye isikhathi, ukugxilwa kwebhafa okukhethiwe kungathathwa njengokufanelekile. Kodwa-ke, ngaphansi kwezimo zokwenyuka kwezinga lokushisa kwemvelo noma lapho usebenzisa amandla kagesi aphezulu, ukuhwamuka kwamanzi ngenxa yokushisa kungadlondlobala, kuphumele ekugxiliseni kwe-buffer ephezulu kakhulu futhi kubangele ngisho nolwelwesi ukuba lome.
Ivolumu yesampula
Ku-cellulose acetate membrane electrophoresis, inani levolumu yesampula linqunywa izici ezihlukahlukene, okuhlanganisa izimo ze-electrophoresis, izakhiwo zesampula ngokwalo, izindlela zokungcolisa, namasu okuthola. Njengomthetho ojwayelekile, uma indlela yokuthola izwela kakhulu, ivolumu yesampula ingaba yincane, okunenzuzo ekuhlukaniseni. Uma ivolumu yesampula idlulele, amaphethini okuhlukaniswa kwe-electrophoretic angase angacaci, futhi ukungcolisa kungase kudle isikhathi. Kodwa-ke, lapho kuhlaziywa ngokobuningi amabhande anamabala ahlukanisiwe kusetshenziswa izindlela zokuthola umbala we-lution, umthamo wesampula akufanele ube mncane kakhulu, njengoba ungabangela amanani aphansi wokumunca wezingxenye ezithile, okuholela kumaphutha aphezulu ekubaleni okuqukethwe kwazo. Ezimweni ezinjalo, umthamo wesampula kufanele ukhuliswe ngendlela efanele.
Ngokuvamile, umthamo wesampula owengezwe kusentimitha ngalinye lomugqa wohlelo lokusebenza lwesampula usukela ku-0.1 kuye ku-5 μL, okulingana nenani lesampula lika-5 kuya ku-1000 μg. Isibonelo, ekuhlaziyweni kwe-serum protein electrophoresis, umthamo wesampula owengezwe kusentimitha ngalinye lomugqa wokufaka isicelo ngokuvamile aweqi ku-1 μL, okulingana no-60 kuya ku-80 μg weprotheni. Kodwa-ke, lapho uhlaziya ama-lipoprotein noma ama-glycoprotein kusetshenziswa indlela efanayo ye-electrophoresis, umthamo wesampula udinga ukwenyuswa ngokuhambisanayo.
Sengiphetha, umthamo wesampula ofaneleka kakhulu kufanele ukhethwe ngokusekelwe ezimweni ezithile ngochungechunge lokuhlolwa kokuqala.
Ukukhethwa kweStaining Solution
Amabhendi ahlukanisiwe ku-cellulose acetate membrane electrophoresis avamise ukuba namabala ngaphambi kokutholwa. Izingxenye zesampula ezihlukene zidinga izindlela ezihlukene zokungcolisa, futhi izindlela zokungcolisa ezifanele i-cellulose acetate membrane electrophoresis zingase zingasebenzi ngokuphelele ephepheni lokuhlunga.
Kunezimiso ezintathu ezinkulu ongakhetha kuzo isixazululo se-stainingi-cellulose acetate membrane. Okokuqala,Odayi abancibilika emanzini kufanele banconywe kunodayi abancibilika ngotshwala ukuze kugwenywe ukushwabana kolwelwesi nokonakala okubangelwa isixazululo esinamabala sakamuva. Ngemuva kokugcoba, kubalulekile ukugeza i-membrane ngamanzi futhi unciphise ubude besikhathi. Uma kungenjalo, i-membrane ingase igoqeke noma incishwe, okungase kuthinte ukutholwa okulandelayo.
Okwesibili, kungcono ukhethe odayi abanokuqina okuqinile kwesampula. Ku-cellulose acetate membrane electrophoresis yamaprotheni e-serum, i-amino black 10B ivame ukusetshenziswa ngenxa yokuhlangana kwayo okunamandla kwezingxenye zeprotheni ze-serum nokuzinza kwayo.
Okwesithathu, kufanele kukhethwe odayi bekhwalithi abathembekile. Abanye odayi, naphezu kokuba negama elifanayo, bangase baqukathe ukungcola okuholela kungemuva elimnyama ngemva kokungcoliswa. Lokhu kungafiphalisa namabhendi ekuqaleni ahlukaniswe kahle, kuwenze kube nzima ukuwahlukanisa.
Okokugcina, ukukhethwa kwesisombululo sokugxila kubalulekile. Ngokwetiyori, kungase kubonakale sengathi ukugxiliswa kwesisombululo esinamabala aphezulu kungaholela ekungcolisweni okuphelele kwezingxenye zesampula kanye nemiphumela engcono yokubola. Nokho, akunjalo. Isibopho esibophezelayo phakathi kwezingxenye zesampula kanye nokudayi kunomkhawulo othile, ongakhuli ngokunyuka kokugxiliswa kwesisombululo sebala. Ngokuphambene nalokho, ukugxila kwesisombululo esinamabala aphezulu ngokweqile akuchithi nje kuphela udayi kodwa futhi kwenza kube nzima ukuthola isizinda esicacile. Ngaphezu kwalokho, lapho ukushuba kombala kufinyelela inani elithile eliphakeme, ijika lokumunca idayi alilandeli ubudlelwano bomugqa, ikakhulukazi ezilinganisweni zobuningi.Ku-cellulose acetate membrane electrophoresis, ukugxiliswa kwesisombululo esinamabala ngokuvamile kuphansi kunalokho okusetshenziswe ku-electrophoresis yephepha.
Imininingwane yokwazi nge-Beijing Liuyi Biotechnology's i-cellulose acetate membraneIthangi le-electrophoresis kanye nesicelo salo se-electrophoresis, sicela uvakashele lapha:
lUkuhlolwa kokuhlukanisa amaprotheni e-serum nge-Cellulose Acetate Membrane
lI-Cellulose Acetate Membrane Electrophoresis
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Ireferensi:I-Electrophoresis(uhlelo lwesibili) kaMnu. Li
Isikhathi sokuthumela: Jun-06-2023