Key Difference – SDS Page vs Native Page
SDS thiab nplooj ntawv ib txwm muaj yog ob hom polyacrylamide gel electrophoresis cov txheej txheem siv hauv Molecular Biology. Qhov sib txawv tseem ceeb ntawm SDS Page thiab Native Page yog hom polyacrylamide gel siv. Nyob rau hauv SDS Page ib denaturing gel yog siv yog li ntawd, molecules raug cais raws li lawv molecular hnyav. Nyob rau hauv sib piv, nyob rau hauv Native Page, uas tsis yog-denaturing gels yog siv. Yog li ntawd, molecules raug cais raws li lawv qhov loj me, nqi thiab cov duab.
Polyacrylamide Gel Electrophoresis (Nplooj) siv cov gel ua los ntawm polymerizing acrylamide monomers nrog methylene bisacrylamide. Cov polyacrylamide yog tawv thiab kub ruaj khov dua li agarose. Cov polyacrylamide gels muaj qhov pore me me uas ua rau muaj kev sib cais ntawm cov protein ntau. Muaj ob hom kev teeb tsa tseem ceeb xws li SDS Page thiab Native Page. SDS Page los yog Sodium-Dodecyl Sulfate Polyacrylamide gel electrophoresis cais cov proteins raws li lawv qhov hnyav molecular. Denaturing gels yog siv hauv SDS Page. Native Page siv cov gels uas tsis yog-denaturing thiab cais cov proteins raws li lawv qhov loj, nqi thiab cov duab (3D conformation).
SDS Page yog dab tsi?
SDS Nplooj ntawv yog cov txheej txheem electrophoretic feem ntau siv los cais cov proteins raws li lawv qhov hnyav molecular. Cov gel yog tsim los ntawm kev ntxiv SDS (Sodium dodecyl sulfate), uas yog cov tshuaj ntxuav tes. SDS dentures proteins rau hauv monomers. SDS yog ib qho tshuaj ntxuav tes anionic. Yog li ntawd, nws ntxiv cov nqi tsis zoo rau cov proteins hauv ntau qhov pH. Thaum lub net tsis zoo nqi yog imparted rau ntawm cov protein molecules, vim hais tias ntawm tus nqi variation, cov complex qauv yog tawg. Vim cov nqi tsis zoo, cov proteins nyiam mus rau qhov kawg. Yog li cov molecules nrog qis molecular hnyav taug kev nrawm dua ntawm cov gel matrix thiab tuaj yeem pom nyob ze ntawm lub anode, thaum cov protein ntau molecular hnyav tau pom ze rau ntawm qhov dej.
Daim duab 01: SDS Page
SDS khi rau cov saw hlau polypeptide yog proportional rau nws cov txheeb ze molecular loj. Yog li, cov molecular loj kuj tuaj yeem txiav txim siab ntawm SDS Page. Kev staining ntawm SDS Page gels yog ua los ntawm bromophenol xiav staining. Daim ntawv thov ntawm SDS nplooj ntawv ntau mus rau qhov loj dua qhov uas nws tuaj yeem siv los kwv yees cov txheeb ze molecular loj thiab txiav txim siab qhov txheeb ze ntawm cov protein ntau hauv cov protein sib xyaw. SDS Page kuj tseem siv tau los txiav txim siab qhov kev faib tawm ntawm cov protein hauv cov protein sib xyaw. SDS Page kuj tseem siv tau los ntxuav thiab tshuaj xyuas cov protein. Nws yog siv los ua cov txheej txheem ua ntej rau sab hnub poob blotting thiab hybridization, uas nyob rau hauv lem yog siv rau protein daim ntawv qhia thiab qhia.
Native Page yog dab tsi?
Native Polyacrylamide gel electrophoresis (Native Page) siv cov gel uas tsis yog-denaturing. Yog li ntawd, SDS los yog lwm yam denaturing tus neeg sawv cev tsis ntxiv rau gel matrix. Hauv Native Page, kev sib cais ntawm cov protein yog nyob ntawm tus nqi thiab qhov loj ntawm cov protein. Yog li, kev txav mus los ntawm cov protein nyob ntawm tus nqi thiab qhov loj ntawm cov protein.
Tus nqi ntawm cov protein nyob ntawm cov saw hlau ntawm cov amino acids. Yog hais tias sab chains raug them tsis zoo, cov protein yuav tau txais tag nrho cov nqi tsis zoo thiab rov ua dua. Proteins khaws 3D conformation vim yog folding uas tshwm sim. Folding tshwm sim los ntawm ntau hom kev cog lus hauv cov protein xws li cov nyiaj disulfide, kev sib cuam tshuam hydrophobic thiab Hydrogen bonds. Yog li ntawd, yog hais tias ib txwm Page yog nqa ntawm ib tug nruab nrab pH, cov proteins yuav raug cais raws li molecular duab ntawm cov protein. Yog li ntawd, Native Page tuaj yeem siv los ua cov txheej txheem rhiab los kuaj xyuas qhov kev hloov pauv lossis kev ua raws li cov protein.
Qhov txiaj ntsig tseem ceeb ntawm nplooj ntawv ib txwm yog tias cov protein siv rau kev txheeb xyuas Nplooj ntawv tuaj yeem rov qab los hauv nws lub xeev qub tom qab Page tsom, vim tias cov protein tsis cuam tshuam thaum lub sijhawm ua haujlwm. Native Page yog cov txheej txheem dhau los ntawm cov txheej txheem siab, thiab kev ruaj ntseg ntawm cov protein ntau ntxiv.
Daim duab 02: Native Page
Thaum ua tiav ntawm cov gel khiav, Native Page gel tuaj yeem pom los ntawm staining nrog bromophenol xiav lossis lwm yam uas tsim nyog staining reagent. Cov ntawv thov ntawm Native Page suav nrog kev sib cais ntawm cov kua qaub nrog rau glycoproteins xws li tib neeg recombinant erythropoietin lossis kev txheeb xyuas cov proteins uas muaj nyob hauv Bovine Serum Albumin (BSA).
Dab tsi zoo sib xws ntawm SDS Page thiab Native Page?
- Ob lub SDS Page thiab Native Page systems siv polyacrylamide gel ua cov matrix ntawm gel.
- Ob leeg siv rau kev sib cais thiab txheeb xyuas cov proteins.
- Ob leeg siv electrophoretic txav mus los cais cov tebchaw.
- Ob leeg tuaj yeem ua tau ntsug lossis kab rov tav (feem ntau ua raws li nplooj ntawv teeb tsa vim tias qhov khiav ntev dua).
- Lub tshuab electrophoresis suav nrog lub gel tank, combs, lub zog yuav tsum tau siv rau kev ua haujlwm ntawm ob txoj kev.
- Kev pom ntawm cov gel tuaj yeem ua tiav los ntawm txoj kev staining hauv ob txoj kev.
Qhov txawv ntawm SDS Page thiab Native Page yog dab tsi?
SDS Page vs Native Page |
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| SDS Page lossis Sodium-dodecyl sulfate Nplooj cais cov proteins raws li lawv qhov hnyav molecular, thiab nws siv cov denaturing gel. | Native Page siv cov gels uas tsis yog-denaturing thiab cais cov proteins raws li lawv qhov loj, nqi thiab cov duab (3D conformation). |
| Hom Gel | |
| A denaturing gel yog siv hauv SDS-nplooj. | A tsis yog-denaturing gel yog siv nyob rau hauv cov nplooj ntawv ib txwm. |
| Kev muaj SDS | |
| SDS tam sim no yog cov tshuaj ntxuav tes kom muab tus nqi tsis zoo ntawm tus qauv hauv SDS nplooj. | SDS tsis muaj nyob hauv nplooj ntawv ib txwm muaj. |
| Sib cais | |
| Kev sib cais ntawm cov protein nyob ntawm qhov hnyav molecular ntawm cov protein hauv SDS nplooj. | Kev sib cais nyob ntawm qhov loj thiab cov duab ntawm cov protein molecule hauv cov nplooj ntawv ib txwm muaj. |
| kev ruaj ntseg ntawm cov protein | |
| Kev ruaj khov ntawm cov protein tsawg hauv SDS nplooj. | Kev ruaj ntseg ntawm cov protein yog siab nyob rau hauv ib txwm nplooj. |
| Rov qab los ntawm Original Protein | |
| Tsis tuaj yeem ua tau raws li nws tau sau tseg hauv SDS nplooj. | Muaj peev xwm ntawm nplooj ntawv ib txwm muaj. |
Summary – SDS Page vs Native Page
SDS Nplooj ntawv thiab Native Page yog ob hom Polyacrylamide gel electrophoresis cov txheej txheem siv los cais cov protein. SDS Nplooj yog kho nrog tshuaj ntxuav tes hu ua SDS. SDS imparts tag nrho cov nqi tsis zoo rau cov protein, uas tom qab ntawd ua rau lub denaturation ntawm cov protein. Yog li ntawd, cov proteins raug cais raws li lawv qhov hnyav molecular. Hauv qhov sib piv, Native Page txheej txheem tsis siv ib tus neeg sawv cev denaturing. Yog li cov proteins yog sib cais raws li lawv qhov loj lossis cov duab. Nov yog qhov sib txawv ntawm SDS nplooj ntawv thiab nplooj ntawv ib txwm.
