Principle
Cellulose acetate film electrophoresis inzira ye electrophoresis inoshandisa cellulose acetate film serutsigiro. Chiitiko umo machaji particles anofamba akananga kune yakatarisana ne electrode pasi pechiito chemunda wemagetsi inonzi electrophoresis. Sezvo puroteni yega yega ine chaiyo isoelectric point, kana puroteni ikaiswa mune mhinduro ine pH yakaderera pane isoelectric point, puroteni inochajirwa zvakanaka uye ichienda kune iyo yakaipa electrode. Pane zvinopesana, inotamira kune yakanaka danda. Nekuti kumhanya kwemapuroteni mamorekuru anofamba mumunda wemagetsi ane chekuita nekuchaja kwavo, chimiro uye saizi yemamorekuru, mapuroteni akasiyana anogona kuparadzaniswa ne electrophoresis. Serum ine akasiyana mapuroteni, ese ane isoelectric point pa pH7.5 kana pasi. Kana serum yaiswa mupH 8.6 buffer yekumhanyisa electrophoresis, ese maprotein eserum anochajiswa zvisina kunaka uye anoenda kudivi rakanaka mundima yemagetsi. Sezvo mapuroteni eSerum akasiyana aine machaji akasiyana papH imwechete, mamolecular particles akasiyana muhukuru, uye nekudaro kumhanya kwekufamba kwakasiyana, uye anoparadzaniswa ne electrophoresis. Iwo isoelectric mapoinzi uye mamorekuru uremu eserum mapuroteni anoratidzwa mutafura pazasi:
| Mapuroteni | Isoelectric points (PI) | Molecular uremu |
| 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 |
Kuedza kuparadzanisa mapuroteni akasiyana muserum yeropa ne cellulose acetate membrane (abbr. CAM) sekutsigira media. CAM imhando yemafuta ane furoefirimu rine yunifomu yakanaka, uye ukobvu i 0.1mm-1.5mm, ine kumwe kutorwa kwemvura.
Zvishandiso, zviridzwa uye reagents zveCAM Electrophoresis
Mienzaniso:serum yeropa remunhu ane hutano
Chiridzwa:magetsi ekupa DYY-6C, electrophoresis tangi DYCP-38C, yepamusoro sampuli kurodha WD-9404
The reagents
1) cellulose acetate membrane 7X9cm
2) PH 8.6 barbitol buffer solution (ionic simba 0.05-0.09, inguva 0.06tid.): tora 1,84g Diethyl barbituric acid, wobva watora 1.03g Diethyl sodium pentobarbital, wedzera imwe mvura yakasvibiswa kupisa kuti iparadze, uye ita. kusvika 1000ml;
3) Stain: Ponceau S 0.2g, Trichloroacetic 3g, wedzera 100ml mvura yakasvibiswa;
4) TBS / T kana PBS / T: 45ml 95% ethanol, 5ml glacial acetic acid, kuwedzera 50ml mvura yakasvibiswa;
5) Kuchenesa Solution: 70ml anhydrous ethanol, 30ml glacial acetic acid.
Experiment Method
1) Gadzirira membrane: Inyura membrane mu barbitol buffer solution 30min-8h, uye ibudise kunze, bvisa imwe mhinduro nepepa rinopinza.
2) Kurodha samples: Siyanisa divi rakakasharara nedivi rakatsetseka re membrane, womaka mutsara uri pa 1.5cm chinhambwe kusvika kumucheto wepamusoro padivi rakashata. Rodha samples nepamusoro sampuli yekurodha chishandiso padivi rakashata. Ongorora: Iwo masampuli anofanirwa kutakurwa padivi rakaomarara re membrane. Mushure mekunge serum samples dzanyatsopinzwa mu membrane, shandura membrane, isa hutsinye rutivi (nemasamples) yakatarisana netangi, uye magumo nemasample anoiswa mune negative electrode.
3) Electrophoresis: Batidza magetsi, 0.4～0.6m A / cm membrane, nguva yekumhanya ndeye 30-45 min. Mushure mekuita electrophoresis, bvisa simba.
4) Kusvibisa uye kuchena: Bvisa membrane kubva mutangi, uye unyudzeit mumushonga wedhayi we 5 min, uye woichenesa mumushonga wekuchenesa nguva nenguva kwe 3-4 nguva kusvikira ruvara rwemashure rwakajeka. Iyo serum mapuroteni inofanira kuratidzwa pamabhandi, uye kazhinji kune nzvimbo shanu, fomu yepamusoro yekupedzisira kumutsara wakaiswa, Albumin, α1-gloulin, α2-gloulin, β-gloulin, γ-gloulin.
5) Kuchengetedza: isa iyo yakaoma elecrtophoresisbandmukuchenesa mhinduro kwemaminetsi gumi-15, wozoitora kunze woinamatira pagirazi rakachena, kana yaoma, inova firimu rinoonekera.band.
ExperimentMhedzisiro
Iyo yekuparadzanisa mhedzisiro yeserum samples yakanaka, uye hapana bhendi tailing phenomenon. Kudzokororwa kwemhedzisiro kunosiyana nekuda kwemaitiro ekuyedza uye nzira dzemuyedzo, uye kudzokorora kwakakwira.
Mhedziso
Iyo yekukurumidza kliniki electrophoresis yekuona system (Electrophoresis tangiDYCP-38C,magetsiDYY-6C uye yepamusoro sampuli kurodha WD-9404) yakagadzirwa neyedukambani Beijing Liuyi Biotechnology Co., Ltdinosangana nezvinodiwa zvekuedza,uyemhedzisiro yacho inogadzirwazve, yakapusa uye inokurumidza, yakakodzerakudzidzisaresearch.
Beijing LiuyiBiotechnology Co., Ltd inoshanda mukugadzira zvigadzirwa zveelecrtophoresis zveindasitiri yesainzi yehupenyu, iyoine nhoroondo yemakore anopfuura makumi mashanu muChina uye kambani inogona kupa zvakagadzikana uye zvemhando yepamusoro zvigadzirwa pasi rese. Kupfuura makore ebudiriro, yakakodzera sarudzo yako!
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Nguva yekutumira: Nov-14-2022