Kev sim rau kev cais cov ntshav protein los ntawm Cellulose Acetate Membrane Electrophoresis

Txoj Cai

Cellulose acetate zaj duab xis electrophoresis yog ib txoj kev electrophoresis uas siv cellulose acetate zaj duab xis los txhawb. Lub phenomenon nyob rau hauv uas them hais mus rau lub opposite electrode nyob rau hauv qhov kev txiav txim ntawm hluav taws xob teb yog hu ua electrophoresis. Txij li txhua cov protein muaj qhov taw qhia tshwj xeeb isoelectric, yog tias cov protein tau muab tso rau hauv cov tshuaj nrog pH qis dua nws qhov isoelectric point, cov protein yuav raug them zoo thiab txav mus rau qhov tsis zoo electrode. Ntawm qhov tsis sib xws, nws txav mus rau tus ncej zoo. Vim tias qhov ceev ntawm cov protein molecules txav mus rau hauv qhov hluav taws xob muaj feem xyuam nrog lawv cov nqi, cov duab thiab qhov loj ntawm cov molecules, cov protein sib txawv tuaj yeem sib cais los ntawm electrophoresis. Serum muaj ntau yam proteins, tag nrho cov uas muaj isoelectric cov ntsiab lus ntawm pH7.5 los yog tsawg dua. Thaum cov ntshav tau muab tso rau hauv pH 8.6 tsis tuaj yeem khiav electrophoresis, tag nrho cov ntshav cov protein yuav raug them tsis zoo thiab yuav txav mus rau qhov zoo hauv qhov hluav taws xob. Txij li thaum muaj ntau yam ntawm cov proteins uas muaj cov nqi sib txawv ntawm tib lub pH, cov khoom molecular yog qhov sib txawv ntawm qhov loj, thiab yog li qhov ceev ntawm kev tsiv teb tsaws txawv, thiab lawv tau sib cais los ntawm electrophoresis. Cov ntsiab lus isoelectric thiab qhov hnyav molecular ntawm cov ntshav cov protein tau qhia hauv cov lus hauv qab no:

Qhov kev sim yog cais cov protein sib txawv hauv cov ntshav ntshav nrog cellulose acetate membrane (abbr. CAM) raws li kev txhawb nqa xov xwm. CAM yog hom foamed loosezaj duab xis nrog zoo sib xws, thiab lub thickness yog 0.1mm-1.5mm, uas muaj ib tug tej yam dej nqus.

Cov ntaub ntawv, cov cuab yeej thiab cov tshuaj reagents rau CAM Electrophoresis

Cov qauv:noj qab nyob zoo tib neeg cov ntshav ntshav

Instrument:fais fab mov DYY-6C, electrophoresis tank DYCP-38C, superior qauv loading WD-9404

Cov reagent

1) cellulose acetate membrane 7X9cm

2) PH 8.6 barbitol buffer tov (ionic zog 0.05-0.09, nws yog 0.06tid lub sij hawm.): noj 1,84g Diethyl barbituric acid, thiab ces noj 1.03g Diethyl sodium pentobarbital, ntxiv ib co distilled dej kom sov kom yaj, thiab ua. mus txog 1000 ml;

3) Stain: Ponceau S 0.2g, Trichloroacetic 3g, ntxiv 100ml distilled dej;

4) TBS / T los yog PBS / T: 45ml 95% ethanol, 5ml glacial acetic acid, ntxiv 50ml distilled dej;

5) Cov tshuaj ntxuav: 70ml anhydrous ethanol, 30ml glacial acetic acid.

Kev sim Method

1) Npaj cov daim nyias nyias: Muab cov membrane tso rau hauv cov tshuaj barbitol buffer 30min-8h, thiab tshem tawm, tshem tawm cov tshuaj ntxiv los ntawm cov ntawv nqus dej.

2) Cov qauv thauj khoom: Paub qhov txawv ntawm qhov ntxhib thiab du ntawm daim nyias nyias, thiab kos ib kab ntawm 1.5cm nrug mus rau sab saum toj kawg ntawm sab ntxhib. Load cov qauv los ntawm superior qauv loading cuab tam nyob rau sab ntxhib. Lus Cim: Cov qauv yuav tsum tau ntim rau ntawm qhov ntxhib ntawm daim nyias nyias. Tom qab cov qauv kuaj ntshav tau nkag mus rau hauv daim nyias nyias, tig dua daim nyias nyias, muab sab ntxhib (nrog cov qauv) ntsej muag rau hauv lub tank, thiab qhov kawg nrog cov qauv muab tso rau hauv qhov tsis zoo electrode.

3) Electrophoresis: Qhib lub hwj huam mov, 0.4～0.6m A / cm membrane, lub sijhawm khiav yog 30-45 min. Tom qab khiav electrophoresis, tua lub hwj chim.

4) Stain thiab huv: Tshem tawm cov membrane ntawm lub tank, thiab rausit rau hauv cov tshuaj zas zas rau 5 feeb, thiab tom qab ntawd ntxuav nws hauv cov tshuaj ntxuav thiab ua 3-4 zaug kom txog thaum cov xim tom qab pom tseeb. Cov proteins hauv cov ntshav yuav tsum tau pom ntawm cov hlua, thiab feem ntau muaj tsib thaj chaw, daim ntawv saum toj kawg nkaus rau kab cim, Albumin, α1-gloulin, α2-gloulin, β-gloulin, γ-gloulin.

5) Preservation: muab cov electophoresis qhuavbandnyob rau hauv cov tshuaj ntxuav rau 10-15 feeb, thiab tom qab ntawd muab nws tawm thiab muab lo rau ntawm lub khob huv, tom qab nws qhuav, nws yuav dhau los ua pob tshab zaj duab xis.band.

Kev simKev tshwm sim

Cov nyhuv sib cais ntawm cov qauv ntshav yog qhov zoo, thiab tsis muaj qhov tshwm sim ntawm qhov tshwm sim. Lub repeatability ntawm cov txiaj ntsig sib txawv vim cov txheej txheem kev sim thiab cov txheej txheem ntawm tus neeg sim, thiab qhov rov ua dua yog siab.

Xaus

Kev soj ntsuam ceev ceev electrophoresis system (Electrophoresis TankDYCP-38C,fais fab movDYY-6C thiab superior qauv loading WD-9404) ua los ntawm pebTuam txhab Beijing Liuyi Biotechnology Co., Ltdua tau raws li qhov xav tau ntawm kev sim,thiabcov txiaj ntsig yog rov tsim dua, yooj yim thiab nrawm, haum raukev qhiakev tshawb fawb.

Beijing LiuyiBiotechnology Co., Ltd tshwj xeeb hauv kev tsim cov khoom lag luam elecrtophoresis rau kev lag luam hauv lub neej, uasmuaj ntau tshaj 50 xyoo keeb kwm nyob rau hauv Suav teb thiab lub tuam txhab muaj peev xwm muab cov khoom ruaj khov thiab cov khoom zoo thoob plaws ntiaj teb. Los ntawm xyoo ntawm txoj kev loj hlob, nws tsim nyog rau koj xaiv!

Peb tam sim no tab tom nrhiav rau cov neeg koom tes, ob qho tib si OEM electrophoresis tank thiab distributors tau txais tos.

Yog xav paub ntxiv txog peb, thov hu rau peb ntawm email[email tiv thaiv]los yog[email tiv thaiv].