Umgaqo
I-cellulose acetate film electrophoresis yindlela ye-electrophoresis usebenzisa ifilimu ye-cellulose acetate njengenkxaso.Isenzeko apho amasuntswana ahlawuliweyo ahambela kwicala le-electrode echaseneyo phantsi kwentshukumo yentsimi yombane ibizwa ngokuba yi-electrophoresis.Ekubeni iprotheni nganye inendawo ethile ye-isoelectric, ukuba iprotheni ifakwe kwisisombululo kunye ne-pH ephantsi kune-isoelectric point, iprotheni iya kuhlawuliswa ngokufanelekileyo kwaye iqhubele kwi-electrode engafanelekanga.Ngokuchasene noko, ihambela kwipali efanelekileyo.Ngenxa yokuba isantya seemolekyuli zeprotheyini ezihamba kwintsimi yombane zihambelana nentlawulo yazo, imilo kunye nobukhulu beemolekyuli, iiprotheni ezahlukeneyo zinokwahlulwa yi-electrophoresis.I-Serum iqulethe iindidi zeeprotheni, zonke zineengongoma ze-isoelectric kwi-pH7.5 okanye ngaphantsi.Xa i-serum ifakwe kwi-pH 8.6 buffer ukuze iqhube i-electrophoresis, zonke iiproteni zeserum zihlawuliswa kakubi kwaye ziya kuhamba kwicala elilungileyo kwindawo yombane.Ekubeni iiprotheni ezahlukeneyo ze-serum zineentlawulo ezahlukeneyo kwi-pH efanayo, iinqununu ze-molecular zihluke ngobukhulu, kwaye ngoko ke isantya sokufuduka sihluke, kwaye sahlulwe yi-electrophoresis.Amanqaku e-isoelectric kunye nobunzima bemolekyuli yeeprotheyini zeserum ziboniswa kwitheyibhile engezantsi:
| Iprotheni | Amanqaku e-Isoelectric (PI) | Ubunzima bemolekyuli |
| Albumin | 4.88 | 69 000 |
| α1-gloulin | 5.00 | 200 000 |
| α2-gloulin | 5.06 | 300 000 |
| β-gloulin | 5.12 | 9 000~150 000 |
| γ-gloulin | 6.85~7.50 | 156 000~ 300 000 |
Uvavanyo kukwahlula iiprotheni ezahlukeneyo kwi-serum yegazi kunye ne-cellulose acetate membrane (abbr. CAM) njengenkxaso yeendaba.I-CAM luhlobo lwee-loos ezinogwebueifilimu kunye neyunifomu elungileyo, kwaye ubukhulu yi-0.1mm-1.5mm, ene-absorption ethile yamanzi.
Izinto eziphathekayo, izixhobo kunye nee-reagents ze-CAM Electrophoresis
Iisampulu:serum yegazi lomntu esempilweni
Isixhobo:amandla ombane i-DYY-6C, itanki ye-electrophoresis DYCP-38C, isampuli ephezulu yokulayisha i-WD-9404
Iireagents
1) cellulose acetate inwebu 7X9cm
2) PH 8.6 isisombululo se-barbitol buffer (amandla e-ionic 0.05-0.09, yi-0.06tid ixesha.): thatha i-1,84g Diethyl barbituric acid, uze uthathe i-1.03g ye-Diethyl sodium pentobarbital, yongeza amanzi adibeneyo ekutshiseni ukunyibilika, kwaye wenze ukuya kuthi ga kwi-1000ml;
3) Ibala: Ponceau S 0.2g, Trichloroacetic 3g, yongeza 100ml amanzi distilled;
4) TBS / T okanye PBS / T: 45ml 95% ethanol, 5ml glacial acetic acid, yongeza 50ml amanzi distilled;
5) Ukucoca Isisombululo: 70ml i-ethanol anhydrous, i-30ml i-acetic acid ye-glacial.
Linge Indlelad
I-1) Lungiselela i-membrane: Gxila i-membrane kwisisombululo se-barbitol buffer 30min-8h, kwaye uyikhuphe, susa isisombululo esongezelelweyo ngephepha le-absorbent.
2) Ukulayisha iisampuli: Ukwahlula icala elibi kunye necala eligudileyo le-membrane, kwaye uphawule umgca kumgama we-1.5cm ukuya ekupheleni okuphezulu kwicala elibi.Layisha iisampulu ngesixhobo sokulayisha isampulu ephezulu kwicala elirhabaxa.Qaphela: Iisampulu kufuneka zilayishwe kwicala elirhabaxa le-membrane.Emva kokuba iisampulu ze-serum zingene ngokupheleleyo kwi-membrane, jika phezu kwenwebu, ubeke icala elirhabaxa (kunye neesampuli) ujonge phantsi kwitanki, kwaye isiphelo ngeesampulu sibekwe kwi-electrode engalunganga.
3) I-Electrophoresis: Vula unikezelo lwamandla, 0.4～I-0.6m A / cm i-membrane, ixesha lokuhamba li-30-45 min.Emva kokusebenza kwe-electrophoresis, cima amandla.
I-4) I-Stain kwaye icoceke: Thatha i-membrane kwi-tank, kwaye ucwiliseit kwisisombululo sedayi kwimizuzu emi-5, uze uhlambulule kwisisombululo sokucoca ngokuphindaphindiweyo ngamaxesha angama-3-4 de kube umbala ongasemva ucacile.Iiprotheni ze-serum kufuneka ziboniswe kwiibhendi, kwaye ngokuqhelekileyo kukho imimandla emihlanu, ifom ephezulu ekupheleni komgca ophawulweyo, i-Albumin, i-α1-gloulin, i-α2-gloulin, i-β-gloulin, i-γ-gloulin.
5) Ukugcinwa: faka i-electrophoresis eyomileyoiqelakwisisombululo sokucoca imizuzu eyi-10-15, kwaye emva koko uyikhuphe kwaye uyibeke kwiglasi ecocekileyo, emva kokuba yomile, iya kuba yifilimu ecacileyo.iqela.
ZamaIsiphumo
Isiphumo sokuhlukana kweesampulu ze-serum zilungile, kwaye akukho nto ye-band ye-tailing.Ukuphindaphindwa kweziphumo kuyahluka ngenxa yeenkqubo zovavanyo kunye neendlela zomfuni, kwaye ukuphindaphinda kuphezulu.
Ukuqukumbela
Inkqubo yokufumanisa i-electrophoresis yeklinikhi ekhawulezayo (Itanki ye-ElectrophoresisI-DYCP-38C,ukunikezwa AmandlaI-DYY-6C kunye nesampula ephezulu yokulayisha i-WD-9404) eveliswe yithuinkampani Beijing Liuyi Biotechnology Co., Ltdiyahlangabezana neemfuno zovavanyo,kwayeiziphumo ziyaphinda-phinda, zilula kwaye zikhawuleza, zifanelekileukufundisauphando.
Beijing LiuyiI-Biotechnology Co., Ltd igxile ekwenzeni iimveliso ze-elecrtophoresis kushishino lwesayensi yobomiinembali engaphezulu kweminyaka engama-50 eTshayina kwaye inkampani inokubonelela ngeemveliso ezizinzileyo nezikumgangatho ophezulu kwihlabathi liphela.Ukutyhubela iminyaka yophuhliso, ilufanele ukhetho lwakho!
Ngoku sikhangela amaqabane, zombini itanki ye-OEM electrophoresis kunye nabasasazi bamkelwe.
Ukuze ufumane inkcazelo engakumbi ngathi, nceda uqhagamshelane nathi nge-imeyile[i-imeyile ekhuselweyo]okanye[i-imeyile ekhuselweyo].
Ixesha lokuposa: Nov-14-2022