Ubushakashatsi bwo gutandukanya poroteyine ya serumu na Cellulose Acetate Membrane Electrophoresis

Ihame

Cellulose acetate ya firime electrophoreis nuburyo bwa electrophoreis ukoresheje firime ya selile ya selile nkinkunga.Ikintu cyashizwemo ibice byerekeza kuri electrode itandukanye nigikorwa cyumuriro w'amashanyarazi cyitwa electrophoreis.Kubera ko buri poroteyine ifite ingingo yihariye ya isoelectric, niba poroteyine ishyizwe mubisubizo hamwe na pH munsi yumwanya wa isoelectric, proteine ​​izishyurwa neza kandi igana kuri electrode mbi.Ibinyuranye, yimukira kuri pole nziza.Kuberako umuvuduko wa molekile ya poroteyine igenda mumashanyarazi ifitanye isano nubushakashatsi bwayo, imiterere nubunini bwa molekile, proteyine zitandukanye zirashobora gutandukanywa na electrophorei.Serumu irimo poroteyine zitandukanye, zose zifite ingingo za isoelectric kuri pH7.5 cyangwa munsi yayo.Iyo serumu ishyizwe muri pH 8.6 kugirango ikore electrophoreis, proteine ​​zose za serumu zishyuzwa nabi kandi zizagenda zerekeza kuruhande rwiza mumashanyarazi.Kubera ko poroteyine zitandukanye za serumu zifite amafaranga atandukanye kuri pH imwe, ibice bya molekile bitandukanye mubunini, bityo umuvuduko wo kwimuka ukaba utandukanye, kandi bigatandukanywa na electrophorei.Ingingo za isoelectric hamwe nuburemere bwa molekuline za poroteyine za serumu bigaragara mu mbonerahamwe ikurikira:

Poroteyine

Ingingo za Isoelectric (PI)

Uburemere bwa molekile

Albumin

4.88

69 000

α1-gloulin

5.00

200 000

α2-gloulin

5.06

300 000

gl-gloulin

5.12

9 000 ~ 150 000

gl-gloulin

6.85 ~ 7.50

156 000 ~ 300 000

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Ubushakashatsi nugutandukanya poroteyine zitandukanye muri serumu yamaraso hamwe na selile selile acetate membrane (abbr. CAM) nkibitangazamakuru byunganira.CAM ni ubwoko bwimyenda myinshiefirime ifite imyenda myiza, kandi ubunini ni 0.1mm-1.5mm, bufite amazi runaka.

Ibikoresho, ibikoresho na reagent ya CAM Electrophoresis

Ingero:ubuzima bwiza bwamaraso yumuntu

Igikoresho:amashanyarazi DYY-6C, ikigega cya electrophoreis DYCP-38C, icyitegererezo cyiza cyo gupakira WD-9404

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Reagents

1) selile ya acetate membrane 7X9cm

2) PH 8.6 ya buffer ya barbitol (imbaraga za ionic 0.05-0.09, ni 0.06tid. kugeza kuri 1000ml;

3) Ikizinga: Ponceau S 0.2g, Trichloroacetic 3g, ongeramo 100ml amazi yatoboye;

4) TBS / T cyangwa PBS / T: 45ml 95% Ethanol, 5ml acide glacial acetike, ongeramo 50ml amazi yatoboye;

5) Igisubizo cyogusukura: 70ml anhydrous ethanol, 30ml acide glacial acetic.

Uburyo bw'igeragezad

1) Tegura ururenda: Shira membrane mumuti wa barbitol buffer 30min-8h, hanyuma uyikuremo, ukureho igisubizo cyinyongera ukoresheje impapuro zinjira.

)Ongeraho icyitegererezo ukoresheje ibikoresho byo hejuru byo gupakira kuruhande.Icyitonderwa: Ingero zigomba gutwarwa kuruhande rwa membrane.Ingero za serumu zimaze kwinjizwa muri membrane, hinduranya membrane, shyira uruhande rutoroshye (hamwe nurugero) rwerekeje hasi kuri tank, hanyuma impera hamwe nicyitegererezo ishyizwe muri electrode mbi.

3) Electrophoresis: Zimya amashanyarazi, 0.40,6m A / cm membrane, igihe cyo gukora ni 30-45 min.Nyuma yo gukora electrophoreis, uzimye amashanyarazi.

4) Siga kandi usukure: Kuramo membrane muri tank, hanyuma wibireit mumuti wo gusiga irangi kuminota 5, hanyuma uyisukure mugisubizo cyogusukura inshuro 3-4 kugeza ibara ryinyuma risobanutse.Poroteyine za serumu zigomba kwerekanwa kuri bande, kandi mubisanzwe hariho zone eshanu, zigakora impera yo hejuru kumurongo wagaragaye, Albumin, α1-gloulin, α2-gloulin, β-gloulin, γ-gloulin.

5) Kubungabunga: shyira elecrtophoresis yumyebandmugusukura igisubizo muminota 10-15, hanyuma ukagikuramo ukagishyira kumirahuri isukuye, namara gukama, bizahinduka firime iboneyeband.

UbushakashatsiIgisubizo

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Ingaruka zo gutandukanya serumu ntangarugero nibyiza, kandi nta bande umurizo.Gusubiramo ibisubizo biratandukanye bitewe nuburyo bwo gukora ibizamini hamwe nuburyo bwabashakashatsi, kandi kubisubiramo ni byinshi.

Umwanzuro

Sisitemu yihuse yubuvuzi bwa electrophoreis (Ikigega cya ElectrophoresisDYCP-38C,amashanyaraziDYY-6C hamwe nicyitegererezo cyo gupakira WD-9404) cyakozwe nuwacuisosiyete Beijing Liuyi Biotechnology Co, Ltd.yujuje ibisabwa mu bushakashatsi,naibisubizo birororoka, byoroshye kandi byihuse, birakwiriyekwigishaubushakashatsi.

Beijing LiuyiBiotechnology Co., Ltd kabuhariwe mu gukora ibicuruzwa bya elecrtophoresis byubuzima bwa siyanse yubuzima, aribyoifite amateka yimyaka irenga 50 mubushinwa kandi isosiyete irashobora gutanga ibicuruzwa bihamye kandi byujuje ubuziranenge kwisi yose.Binyuze mumyaka yiterambere, birakwiye guhitamo!

Ubu turimo gushakisha abafatanyabikorwa, ikigega cya OEM electrophoresis hamwe nababitanga barahawe ikaze.

Kubindi bisobanuro kuri twe, nyamuneka twandikire ukoresheje imeri[imeri irinzwe]cyangwa[imeri irinzwe].


Igihe cyo kohereza: Ugushyingo-14-2022