Koj tuaj yeem tshem koj cov qauv los ntawm ultrafiltration mus rau 1 feem kaum ntawm cov ntim dhau los, uas diluted dua thiab rov ua UF kauj ruam. Txoj kev zoo tshaj plaws rau des alting yog des alting los ntawm gel pom tawm tsam 50 mM tsis, dua li ultrafiltration (classical, tsis yog centrifugal).
Dab tsi yog qhov kev tshem tawm ntawm cov protein purification?
Des alting yog siv txhawm rau tshem cov ntsev los ntawm cov tshuaj protein, phenol lossis nucleotides tsis suav nrog los ntawm nucleic acids lossis ntau dhau kev sib txuas lossis sau cov tshuaj reagents los ntawm cov protein sib txuas.
Koj ua li cas ntxuav cov protein?
Nyob rau hauv kev ua kom huv ntawm cov protein ntau, thawj kauj ruam ntawm kev cais cov proteins yog precipitation nrog ammonium sulfate (NH 4) 2SO4. Qhov no yog ua los ntawm kev ntxiv ntau ntxiv ntawm ammonium sulfate thiab sau cov feem sib txawv ntawm precipitated protein. Tom qab ntawd, ammonium sulfate tuaj yeem raug tshem tawm los ntawm kev lim ntshav.
Koj ua li cas tsis pauv cov proteins?
Buffer exchange, txawm li cas los xij, yog ua los ntawm thawj qhov sib npaug nrog cov kab resin nrog cov buffer tus qauv yuav tsum xaus rau hauv. lub kem yuav raug hloov los ntawm cov tshuaj uas kem yog pre-equilibrated.
Qhov twg ntawm cov txheej txheem chromatography hauv qab no tuaj yeem siv rau kev tshem tawm cov protein?
Raws li tau hais los saum no, gel pom tuaj yeem siv tau zoo rau cov proteindes alting thiab yog ua tiav los ntawm thawj equilibrating gel filtration kem nrog dej. Txawm li cas los xij, kev sib pauv tsis yog ua tiav los ntawm thawj qhov sib npaug ntawm kem resin nrog lub hom phiaj tsis.
