Isimiso
I-cellulose acetate film electrophoresis iyindlela ye-electrophoresis esebenzisa ifilimu ye-cellulose acetate njengendlela yokusekela.Isenzakalo lapho izinhlayiya ezikhokhisiwe zihamba zibheke kwelinye i-electrode ngaphansi kwesenzo sensimu kagesi ibizwa ngokuthi i-electrophoresis.Njengoba iphrotheni ngayinye inephuzu elithile le-isoelectric, uma iphrotheni ibekwe esixazululweni esine-pH ephansi kunephoyinti layo le-isoelectric, iphrotheni izoshajwa kahle futhi iqonde ku-electrode eyinegethivu.Ngokuphambene nalokho, iya esigxotsheni esihle.Ngenxa yokuthi ijubane lama-molecule amaprotheni ahamba endaweni kagesi lihlobene nokushaja kwawo, ukuma nobukhulu bama-molecule, amaprotheni ahlukene angahlukaniswa yi-electrophoresis.ISerum iqukethe amaprotheni ahlukahlukene, wonke anamaphuzu e-isoelectric ku-pH7.5 noma ngaphansi.Uma i-serum ibekwe ku-pH 8.6 buffer ukuze isebenze i-electrophoresis, wonke amaprotheni e-serum ashajwa kabi futhi azoya ohlangothini oluhle endaweni kagesi.Njengoba amaprotheni e-serum ahlukahlukene anezindleko ezihlukene nge-pH efanayo, izinhlayiya zamangqamuzana zihlukile ngosayizi, futhi ngaleyo ndlela ijubane lokufuduka lihlukile, futhi lihlukaniswa yi-electrophoresis.Amaphuzu we-isoelectric kanye nezisindo zamangqamuzana amaprotheni e-serum aboniswa etafuleni elingezansi:
| Amaprotheni | Amaphoyinti e-Isoelectric(PI) | Isisindo samangqamuzana |
| I-albhamuin | 4.88 | 69 000 |
| α1-gloulin | 5.00 | 200 000 |
| α2-gloulin | 5.06 | 300 000 |
| i-β-gloulin | 5.12 | 9 000 ~ 150 000 |
| γ-gloulin | 6.85~7.50 | 156 000 ~ 300 000 |
Ukuhlolwa okokwehlukanisa amaprotheni ahlukene ku-serum yegazi nge-cellulose acetate membrane (abbr. CAM) njengemidiya yokusekela.I-CAM iwuhlobo lwama-loos anegwebueifilimu enomfaniswano omuhle, futhi ukujiya kungu-0.1mm-1.5mm, okunokumuncwa okuthile kwamanzi.
Izinto, amathuluzi kanye nama-reagents we-CAM Electrophoresis
Amasampuli:i-serum yegazi lomuntu enempilo
Insimbi:ukunikezwa kwamandla kwe-DYY-6C, ithangi le-electrophoresis i-DYCP-38C, isampula ephakeme yokulayisha i-WD-9404
Ama-reagents
1) i-cellulose acetate membrane 7X9cm
2) PH 8.6 isixazululo sebhafa ye-barbitol (amandla e-ionic 0.05-0.09, isikhathi esingu-0.06tid.): thatha u-1,84g we-Diethyl barbituric acid, bese uthatha u-1.03g we-Diethyl sodium pentobarbital, engeza amanzi ancibilikisiwe ukuze ushise ukuze uncibilike, bese wenza kuze kufike ku-1000 ml;
3) Ibala: Ponceau S 0.2g, Trichloroacetic 3g, engeza 100ml amanzi distilled;
4) TBS/T noma PBS/T: 45ml 95% ethanol, 5ml glacial acetic acid, engeza 50ml amanzi distilled;
I-5) Isixazululo Sokuhlanza: 70ml i-ethanol anhydrous, 30ml i-glacial acetic acid.
Indlela Yokuhlolad
1) Lungisa ulwelwesi: Gxilisa ulwelwesi kusixazululo sebhafa ye-barbitol 30min-8h, bese uyikhipha, susa isixazululo esengeziwe ngephepha elimuncayo.
2) Amasampula okulayisha: Hlukanisa uhlangothi olumaholoholo nohlangothi olubushelelezi lwelwelwesi, bese umaka umugqa osebangeni elingu-1.5cm ukuya ekugcineni ongaphezulu ohlangothini olumaholoholo.Layisha amasampuli ngethuluzi lokulayisha lesampula eliphakeme ohlangothini olumaholo.Qaphela: Amasampula kufanele alayishwe ohlangothini olunzima lwe-membrane.Ngemva kokuba amasampula e-serum esengene ngokugcwele kulwelwesi, vula ulwelwesi, ubeke uhlangothi olumahhadlahhadla (namasampula) lubheke phansi ethangini, bese ekugcineni enamasampuli kufakwe ku-electrode engalungile.
3) I-Electrophoresis: Vula ukunikezwa kwamandla, 0.4～I-membrane engu-0.6m A/cm, isikhathi sokusebenza siyi-30-45 min.Ngemuva kokusebenzisa i-electrophoresis, cisha amandla.
4) Gcoba futhi uhlanze: Khipha ulwelwesi ethangini, bese ucwilisait ungene kwisisombululo sikadayi imizuzu emi-5, bese uyihlanza kusixazululo sokuhlanza ngokuphindaphindiwe izikhathi ezingu-3-4 kuze kube yilapho umbala wangemuva ucacile.Amaprotheni e-serum kufanele aboniswe kuma-bands, futhi ngokuvamile kunezindawo ezinhlanu, ifomu eliphezulu ekugcineni emgqeni omakiwe, i-Albumin, i-α1-gloulin, i-α2-gloulin, i-β-gloulin, i-γ-gloulin.
5) Ukulondolozwa: faka i-elecrtophoresis eyomileibhandeekuhlanzeni isisombululo imizuzu eyi-10-15, bese uyikhipha bese uyinamathisela engilazini ehlanzekile, ngemuva kokoma, izoba ifilimu esobala.ibhande.
IsivivinyoUmphumela
Umthelela wokuhlukanisa wamasampula e-serum muhle, futhi akukho mkhuba wokulandelanisa kwebhande.Ukuphindaphinda kwemiphumela kuyehluka ngenxa yezinqubo zokuhlola kanye nezindlela zomhloli, futhi ukuphindaphinda kuphezulu.
Isiphetho
Uhlelo lokuthola i-electrophoresis olusheshayo emitholampilo (I-Electrophoresis tankI-DYCP-38C,ukunikezwa kwamandla kagesiI-DYY-6C kanye nesampula ephezulu yokulayisha i-WD-9404) ekhiqizwe ngabethuinkampani Beijing Liuyi Biotechnology Co., Ltdihlangabezana nezidingo zokuhlola,futhiimiphumela iyaphindaphindeka, ilula futhi iyashesha, ifanelekileukufundisaucwaningo.
Beijing LiuyiI-Biotechnology Co., Ltd igxile ekukhiqizeni imikhiqizo ye-elecrtophoresis embonini yesayensi yempilo, okuyintoinomlando weminyaka engaphezu kwengama-50 eChina futhi inkampani ingahlinzeka ngemikhiqizo ezinzile nesezingeni eliphezulu emhlabeni wonke.Ngeminyaka yokuthuthuka, kufanelekile ukukhetha kwakho!
Manje sifuna ozakwethu, kokubili ithangi le-OEM electrophoresis kanye nabasabalalisi bamukelekile.
Ukuze uthole ukwaziswa okwengeziwe ngathi, sicela usithinte nge-imeyili[i-imeyili ivikelwe]noma[i-imeyili ivikelwe].
Isikhathi sokuthumela: Nov-14-2022