Sabelana ngeengqwalaselo ezininzi kwiveki ephelileyo ngokusebenzisa i-cellulose acetate membrane electrophoresis, kwaye siyakugqibezela esi sihloko apha namhlanje ngereferensi yakho.
Ukukhethwa kwe Ugxininiso lwe-Buffer
I-concentration ye-buffer esetyenziswe kwi-cellulose i-acetate membrane i-electrophoresis ingaphantsi ngokubanzi kuneyo isetyenziswe kwi-electrophoresis yamaphepha.I-pH esetyenziswa rhoqo yi-8.6Bi-arbital buffer idla ngokukhethwa phakathi koluhlu lwe-0.05 mol/L ukuya kwi-0.09 mol/L.Xa ukhetha ugxininiso, ukuzimisela kwangaphambili kwenziwa.Ngokomzekelo, ukuba ubude be-membrane strip phakathi kwe-electrodes kwigumbi le-electrophoresis yi-8-10cm, i-voltage ye-25V iyadingeka ngesentimitha yobude be-membrane ubude, kwaye ukuqina kwangoku kufuneka kube yi-0.4-0.5 mA ngesentimitha yobubanzi be-membrane.Ukuba ezi xabiso azifezekanga okanye zigqithiswa ngexesha le-electrophoresis, i-concentration ye-buffer kufuneka inyuswe okanye ihlanjululwe.
Ukugxininiswa kwe-buffer ephantsi kakhulu kuya kubangela intshukumo ekhawulezayo yeebhendi kunye nokwanda kobubanzi bebhendi.Kwelinye icala, ugxininiso lwe-buffer oluphezulu kakhulu luya kuthoba isantya sokufuduka kwebhendi, kwenze kube nzima ukwahlula amaqela athile okwahlula.
Kufuneka kuqatshelwe ukuba kwi-cellulose i-acetate membrane electrophoresis, inxalenye ebalulekileyo yangoku iqhutywe ngesampuli, eyenza ubushushu obuninzi.Ngamanye amaxesha, ugxininiso olukhethiweyo lwe-buffer lunokubonwa lufanelekile.Nangona kunjalo, phantsi kweemeko zokonyuka kobushushu bendalo okanye xa kusetyenziswa amandla ombane aphezulu, ukuphuphuma kwamanzi ngenxa yobushushu kunokuqina, okukhokelela kugxininiso oluphezulu kakhulu lwe-buffer kwaye lude lubangele ukuba inwebu yome.
Umthamo wesampuli
Kwi-cellulose acetate membrane electrophoresis, ubungakanani besampulu yesampulu imiselwa zizinto ezahlukeneyo, kubandakanya iimeko ze-electrophoresis, iipropathi zesampulu ngokwayo, iindlela zokungcolisa, kunye nobuchule bokubona.Njengomgaqo jikelele, okukhona uvakalelo lwendlela yokubona, incinci ivolumu yesampulu inokuba yinto eluncedo kukwahlukana.Ukuba umthamo wesampulu ugqithise, iipateni zokwahlula i-electrophoretic zisenokungacaci, kwaye ukungcolisa kungadla ixesha.Nangona kunjalo, xa uhlalutya ngokobungakanani iibhendi ezigudisiweyo ezahluliweyo zisebenzisa iindlela zokubona umbala we-lution, umthamo wesampulu akufanele ube mncinci kakhulu, njengoko unokubangela amaxabiso asezantsi okufunxa kumacandelo athile, okukhokelela kwiimpazamo eziphezulu ekubaleni umxholo wabo.Kwiimeko ezinjalo, umthamo wesampuli kufuneka unyuswe ngokufanelekileyo.
Ngokuqhelekileyo, umthamo wesampuli wongezwa kwisentimitha nganye yomgca wesicelo sesampuli ukusuka kwi-0.1 ukuya kwi-5 μL, ilingana nesamba sesampuli ye-5 ukuya kwi-1000 μg.Ngokomzekelo, kwi-protein ye-serum ye-protein ye-electrophoresis uhlalutyo, umthamo wesampulu eyongeziweyo kwi-centimeter nganye yomgca wesicelo ngokubanzi awudluli i-1 μL, ilingana ne-60 ukuya kwi-80 μg yeprotheni.Nangona kunjalo, xa uhlalutya i-lipoprotein okanye i-glycoprotein usebenzisa indlela ye-electrophoresis efanayo, umthamo wesampulu kufuneka unyuswe ngokuhambelanayo.
Ukuqukumbela, owona mthamo ufanelekileyo wesampulu kufuneka ukhethwe ngokusekelwe kwiimeko ezithile ngothotho lweemvavanyo zangaphambili.
Ukukhethwa kweSisombululo sokuStaining
Iibhanti ezahluliweyo kwi-cellulose i-acetate membrane electrophoresis zihlala zingcoliswa ngaphambi kokuba zibhaqwe.Amacandelo ahlukeneyo esampulu afuna iindlela ezahlukeneyo zokucoca, kwaye iindlela zokucoca ezifanelekileyo kwi-cellulose i-acetate membrane electrophoresis ayinakusetyenziswa ngokupheleleyo kwiphepha lokucoca.
Kukho imigaqo emithathu engundoqo yokukhetha isisombululo sokucocai-cellulose acetate inwebu.Okokuqala,Idayi ezinyibilikayo zamanzi kufuneka zikhethwe ngaphezu kwedayi ezinyibilikayo ngotywala ukuze zigweme i-membrane shrinkage kunye ne-deformation ebangelwa yisisombululo se-staining yokugqibela.Emva kokungcola, kubalulekile ukuhlamba i-membrane ngamanzi kwaye unciphise ubude bexesha.Ngaphandle koko, i-membrane inokugobeka okanye ishwabene, nto leyo enokuchaphazela ukubhaqwa okulandelayo.
Okwesibini, kuyakhethwa ukuba ukhethe iidayi ezinokuqina okuqinileyo kwisampulu.Kwi-cellulose i-acetate membrane electrophoresis yeeprotheni ze-serum, i-amino emnyama i-10B isetyenziswa ngokuqhelekileyo ngenxa ye-staining affinity eyomeleleyo ye-serum protein components kunye nokuzinza kwayo.
Okwesithathu, iidayi ezisemgangathweni kufuneka zikhethwe.Ezinye iidayi, nangona zinegama elifanayo, zinokuqulatha ukungcola okukhokelela kwimvelaphi emnyama emva kokungcola.Oku kunokude kufiphale amabhanti aqale ahlulwe kakuhle, kuwenza kube nzima ukuwahlula.
Okokugqibela, ukhetho lokuxinana kwesisombululo lubalulekile.Ngokwethiyori, kunokubonakala ngathi ugxininiso oluphezulu lwesisombululo lunokukhokelela ekungcolisweni okucokisekileyo kwamacandelo esampulu kunye neziphumo ezingcono zokungcolisa.Nangona kunjalo, oku akunjalo.Ubudlelwane obubophelelayo phakathi kwamacandelo esampuli kunye nedayi inomda othile, onganyuki ngokunyuka koxinzelelo lwesisombululo.Ngokuchasene noko, i-concentration yesisombululo esiphezulu esigqithisileyo ayichithi nje kuphela idayi kodwa yenza kube nzima ukufezekisa imvelaphi ecacileyo.Ngaphezu koko, xa ubukhulu bombala bufikelela kwixabiso elithile eliphezulu, i-absorbection curve yedayi ayilandeli ubudlelwane bomgca, ngokukodwa kwimilinganiselo yobungakanani.Kwi-cellulose acetate membrane electrophoresis, i-concentration ye-staining solution ingaphantsi kuneyo isetyenziswe kwi-electrophoresis yephepha.
Iinkcukacha ukwazi malunga neBeijing Liuyi Biotechnology's i-cellulose acetate inwebuItanki ye-electrophoresis kunye nesicelo sayo se-electrophoresis, nceda undwendwele apha:
lUlingo lokwahlula iprotein ye-serum nge-Cellulose Acetate Membrane
lI-Cellulose Acetate Membrane Electrophoresis
Ukuba unayo nayiphi na isicwangciso sokuthenga iimveliso zethu, nceda ungalibazisi ukudibana nathi.Ungasithumela umyalezo nge-imeyile[i-imeyile ekhuselweyo]okanye[i-imeyile ekhuselweyo], okanye nceda usitsalele umnxeba +86 15810650221 okanye wongeze iWhatsapp +86 15810650221, okanye Wechat: 15810650221.
Isalathiso:Electrophoresis(uhlelo lwesibini) nguMnu. Li
Ixesha lokuposa: Jun-06-2023