Kumukānāwai
ʻO ka cellulose acetate film electrophoresis kahi hana electrophoresis e hoʻohana ana i kahi kiʻi cellulose acetate ma ke ʻano he kākoʻo. ʻO ke ʻano o ka neʻe ʻana o nā ʻāpana i hoʻopaʻa ʻia i ka electrode ʻē aʻe ma lalo o ka hana o ke kahua uila i kapa ʻia ʻo electrophoresis. No ka mea, loaʻa i kēlā me kēia protein kahi helu isoelectric kikoʻī, inā hoʻokomo ʻia kahi protein i loko o kahi hopena me ka pH haʻahaʻa ma mua o kona helu isoelectric, e hoʻopiʻi maikaʻi ʻia ka protein a neʻe i ka electrode maikaʻi ʻole. ʻO ka mea ʻē aʻe, neʻe ia i ka pou maikaʻi. No ka mea e pili ana ka wikiwiki o nā molekele protein e neʻe ana i loko o ke kahua uila i kā lākou uku, ke ʻano a me ka nui o nā molekala, hiki ke hoʻokaʻawale ʻia nā protein like ʻole e ka electrophoresis. Loaʻa i ka Serum nā ʻano protein, ʻo ia mau mea āpau i nā helu isoelectric ma ka pH7.5 a i ʻole. Ke waiho ʻia ka serum i loko o ka pH 8.6 buffer e holo i ka electrophoresis, ua hoʻopiʻi ʻia nā protein serum a pau a neʻe i ka ʻaoʻao maikaʻi o ke kahua uila. No ka mea he ʻokoʻa nā koina serum like ʻole i ka pH like, ʻokoʻa ka nui o nā ʻāpana molekala, a no laila ʻokoʻa ka wikiwiki o ka neʻe ʻana, a ua hoʻokaʻawale ʻia lākou e ka electrophoresis. Hōʻike ʻia nā helu isoelectric a me nā paona molekala o nā protein serum ma ka papa ma lalo:
| Polokina | Nā helu Isoelectric (PI) | Kaumaha molekula |
| Albumina | 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 |
ʻO ka hoʻokolohua e hoʻokaʻawale i nā protein like ʻole i loko o ka serum koko me ka cellulose acetate membrane (abbr. CAM) ma ke ʻano he media kākoʻo. ʻO ka CAM kahi ʻano o nā loos foamedeʻO ke kiʻiʻoniʻoni me ka ʻaʻahu maikaʻi, a ʻo ka mānoanoa he 0.1mm-1.5mm, aia kekahi wai absorption.
Nā mea, nā mea hana a me nā reagents no CAM Electrophoresis
Nā laʻana:serum koko kanaka olakino
Mea hana:lako mana DYY-6C, pahu electrophoresis DYCP-38C, hoʻouka laʻana kiʻekiʻe WD-9404
ʻO nā reagents
1) cellulose acetate membrane 7X9cm
2) PH 8.6 barbitol buffer solution (ionic ikaika 0.05-0.09, he 0.06tid ka manawa.): lawe i ka 1,84g Diethyl barbituric acid, a laila lawe i ka 1.03g Diethyl sodium pentobarbital, e hoʻohui i kekahi wai distilled e wela e hoʻoheheʻe, a hana. a hiki i 1000ml;
3) Pahu: ʻO Ponceau S 0.2g, Trichloroacetic 3g, hoʻohui i 100ml wai i hoʻoheheʻe ʻia;
4) TBS / T a i ʻole PBS / T: 45ml 95% ethanol, 5ml glacial acetic acid, hoʻohui i 50ml wai i hoʻoheheʻe ʻia;
5) Mea Hoʻomaʻemaʻe: 70ml anhydrous ethanol, 30ml glacial acetic acid.
Hoʻāʻo Method
1) E hoʻomākaukau i ka membrane: E hoʻokomo i ka membrane i loko o ka barbitol buffer solution 30min-8h, a lawe i waho, e hoʻoneʻe i ka hopena hou ma ka pepa absorbent.
2) Hoʻouka ʻana i nā laʻana: E hoʻokaʻawale i ka ʻaoʻao ʻōniʻoniʻo a me ka ʻaoʻao maʻemaʻe o ka membrane, a e kaha i kahi laina ma 1.5cm mamao a hiki i ka ʻaoʻao luna ma ka ʻaoʻao ʻala. E hoʻouka i nā laʻana ma o ka mea hana hoʻouka hāpana kiʻekiʻe ma ka ʻaoʻao ʻala. 'Ōlelo Aʻo: Pono e hoʻouka ʻia nā laʻana ma ka ʻaoʻao ʻala o ka membrane. Ma hope o ka hoʻokomo piha ʻana o ka serum i loko o ka membrane, e hoʻohuli i ka membrane, e kau i ka ʻaoʻao ʻōpala (me nā laʻana) i lalo i ka pahu, a waiho ʻia ka hopena me nā laʻana i ka electrode maikaʻi ʻole.
3) Electrophoresis: E ho'āla i ka mana, 0.4～0.6m A / cm membrane, ʻo ka manawa holo he 30-45 min. Ma hope o ka holo electrophoresis, hoʻopau i ka mana.
4) Pahu a hoʻomaʻemaʻe: Wehe i ka membrane mai ka pahu, a hoʻoinuit i loko o ka wai waihoʻoluʻu no 5 min, a laila e hoʻomaʻemaʻe i loko o ka hoʻomaʻemaʻe hoʻomaʻemaʻe pinepine no 3-4 mau manawa a maopopo ke kala hope. Pono e hōʻike ʻia nā protein serum ma nā kaula, a ma ka maʻamau he ʻelima mau ʻāpana, e hoʻokumu i ka hope o ka laina i kaha ʻia, Albumin, α1-gloulin, α2-gloulin, β-gloulin, γ-gloulin.
5) Mālama: kau i ka electrophoresis maloʻohuii ka hoʻomaʻemaʻe hoʻomaʻemaʻe no 10-15 mau minuke, a laila e lawe i waho a hoʻopaʻa iā ia ma ke aniani maʻemaʻe, ma hope o ka maloʻo ʻana, e lilo ia i kiʻi ʻoniʻoni.hui.
HoaoKa hopena
Maikaʻi ka hopena hoʻokaʻawale o nā laʻana serum, a ʻaʻohe mea hōʻikeʻike band tailing. ʻOkoʻa ka hoʻihoʻi hou ʻana o nā hopena ma muli o nā kaʻina hana hoʻāʻo a me nā ʻano o ka mea hoʻokolohua, a ʻoi aku ka kiʻekiʻe o ka repeatability.
Ka hopena
ʻO ka ʻōnaehana ʻike electrophoresis wikiwiki (pahu ElectrophoresisDYCP-38C,lako ikehuDYY-6C a me ka laʻana kiʻekiʻe e hoʻouka ana i ka WD-9404) i hana ʻia e kā mākouhui ʻo Beijing Liuyi Biotechnology Co., Ltdhoʻokō i nā koi hoʻokolohua,ahiki ke hana hou ʻia nā hopena, maʻalahi a wikiwiki hoʻi, kūpono noao ananoiʻi.
Pekina LiuyiʻO Biotechnology Co., Ltd ka mea kūikawā i ka hana ʻana i nā huahana electrtophoresis no ka ʻoihana ʻepekema ola.ʻoi aku ka nui o 50 mau makahiki ma Kina a hiki i ka hui ke hāʻawi i nā huahana kūpaʻa a kiʻekiʻe a puni ka honua. Ma o nā makahiki o ka hoʻomohala ʻana, kūpono ia i kāu koho!
Ke ʻimi nei mākou i nā hoa, ʻelua OEM electrophoresis tank a me nā mea hoʻolaha i hoʻokipa ʻia.
No ka ʻike hou aku e pili ana iā mākou, e ʻoluʻolu e kelepona mai iā mākou ma ka leka uila[palapala leka uila]a i ʻole[palapala leka uila].
Ka manawa hoʻouna: Nov-14-2022