Qhov sib txawv tseem ceeb ntawm chromatofocusing thiab isoelectric focusing yog tias chromatofocusing yog ib hom kab chromatography txoj kev uas siv ion exchange resins thaum isoelectric focusing yog hom electrophoresis cov txheej txheem uas siv immobilized pH gradient gels.
Chromatofocusing thiab isoelectric focusing yog ob txoj kev uas cais cov proteins raws li lawv cov ntsiab lus isoelectric. Ob qho tib si chromatofocusing thiab isoelectric focusing muaj zog daws teeb meem zoo. Chromatofocusing yog ib qho kev daws teeb meem siab kem chromatography txoj kev thaum isoelectric focusing yog ib txoj kev sib cais electrophoretic. Ntxiv mus, chromatofocusing tsis siv hluav taws xob teb thaum isoelectric tsom xav tau hluav taws xob teb.
Chromatofocusing yog dab tsi?
Chromatofocusing yog hom kev ua haujlwm siab chromatography uas cais cov protein raws li qhov sib txawv ntawm lawv qhov isoelectric point. Nws siv cov ion txauv resin kem thiab ib qho kev tsim pH gradient sab hauv. Tsis zoo li isoelectric focusing, chromatofocusing tsis koom nrog hluav taws xob teb. Chromatofocusing yog cov txheej txheem purification muaj zog uas tuaj yeem daws cov molecules zoo sib xws txawm tias tsuas yog txawv ntawm 0.02 pH units. Yog li ntawd, chromatofocusing yog qhov zoo tshaj plaws raws li qhov kawg polishing kauj ruam thaum xav tau kev daws teeb meem siab hauv kev sib cais cov khoom zoo sib xws.
Nyob rau hauv chromatofocusing, tus qauv yog siv rau hauv kem los ntawm kev sib xyaw nrog qhov pib tsis. Cov proteins uas nyob rau ntawm pH saum toj no lawv cov isoelectric point yog qhov tsis zoo thiab lawv nyob ze rau sab saum toj ntawm kem thaum lwm cov proteins uas nyob rau ntawm pH hauv qab lawv qhov isoelectric point, tsiv mus rau sab. Cov protein nrog qhov siab tshaj plaws isoelectric point yog eluted ua ntej thaum cov protein nrog qhov qis tshaj isoelectric yog eluted kawg ntawm kab. Cov txheej txheem no cuam tshuam los ntawm ntau yam xws li nag lossis daus protein thiab linearity ntawm pH gradient.
Isoelectric Focusing yog dab tsi?
Isoelectric focusing yog ib qho txheej txheem uas cais cov molecules sib txawv raws li lawv qhov isoelectric point. Nws tseem hu ua electrofocusing. Cov txheej txheem no feem ntau yog siv rau kev sib cais cov protein hauv ib gel. Hauv isoelectric tsom, cov qauv yog ntxiv rau hauv immobilized pH gradient (IPG) gels. IPG gel yog acrylamide gel matrix copolymerized nrog pH gradient. Cov proteins tsiv mus rau cathode kom txog thaum lawv pom thaj tsam pH ntawm lawv cov ntsiab lus isoelectric. Thaum cov protein nce mus txog pH uas tsis muaj nuj nqis, kev tsiv teb tsaws chaw nres thiab muab ib qho chaw ruaj khov. Ib yam li ntawd, txhua cov protein muab ib pawg ntawm qhov taw tes hauv pH gradient sib haum rau nws qhov isoelectric point.
Daim duab 01: Isoelectric Focusing
Isoelectric tsom xam yog thawj kauj ruam ntawm ob-dimensional gel electrophoresis. Hauv cov txheej txheem ntawd, cov proteins yog thawj zaug sib cais raws li lawv cov ntsiab lus isoelectric los ntawm isoelectric tsom thiab tom qab ntawd sib cais los ntawm lawv qhov hnyav molecular los ntawm SDS-PAGE.
Dab tsi yog qhov zoo sib xws ntawm Chromatofocusing thiab Isoelectric Focusing?
- Chromatofocusing thiab isoelectric focusing yog ob txoj kev daws teeb meem.
- Ob leeg yog raws li isoelectric point ntawm cov proteins.
- Cov tswv yim no siv pH gradient.
- Ob txoj haujlwm yog qhia rau tus yam ntxwv ntawm tib neeg hemoglobin variants.
- Lawv tuaj yeem daws cov molecules qhov twg pI qhov tseem ceeb sib txawv los ntawm tsawg li 0.02 pH units.
Qhov txawv ntawm Chromatofocusing thiab Isoelectric Focusing yog dab tsi?
Chromatofocusing yog ib qho kev daws teeb meem ntawm kab chromatography txheej txheem uas cais cov proteins raws li lawv qhov isoelectric point. Hauv qhov sib piv, isoelectric focusing yog cov txheej txheem electrophoretic uas cais cov proteins raws li lawv qhov isoelectric point los ntawm kev siv cov hluav taws xob raws li pH gradient tsim nyob rau hauv capillary. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm chromatofocusing thiab isoelectric focusing.
Lwm qhov txawv ntawm chromatofocusing thiab isoelectric focusing yog tias chromatofocusing tsis koom nrog hluav taws xob thaum isoelectric tsom xav tau hluav taws xob teb.
Summary – Chromatofocusing vs Isoelectric Focusing
Chromatofocusing thiab isoelectric focusing yog ob txoj kev zoo sib xws uas muaj peev xwm daws tau cov molecules siab, tshwj xeeb tshaj yog cov molecules zoo sib xws. Chromatofocusing yog ib qho kev daws teeb meem siab kab chromatography txheej txheem thaum isoelectric focusing yog ib txoj kev electrophoresis. Chromatofocusing tsis koom nrog qhov hluav taws xob, tsis zoo li isoelectric focusing. Yog li, qhov no qhia txog qhov sib txawv ntawm chromatofocusing thiab isoelectric focusing.
