Qhov Tseem Ceeb Qhov txawv - SDS Page vs Western Blot
Western blotting yog cov txheej txheem uas kuaj pom cov protein tshwj xeeb los ntawm cov qauv protein. Cov txheej txheem no yog ua los ntawm ob peb kauj ruam tseem ceeb: gel electrophoresis, blotting, thiab hybridization. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS Page) yog ib hom gel electrophoresis txheej txheem uas yog siv los cais cov proteins raws li lawv qhov ntau thiab tsawg (qhov hnyav molecular). Western blot yog ib daim ntawv tshwj xeeb ntawm daim nyias nyias uas siv los hloov tib cov qauv ntawm cov proteins hauv SDS Page. Qhov sib txawv tseem ceeb ntawm SDS Page thiab western blot yog SDS Page tso cai rau kev sib cais ntawm cov proteins hauv kev sib tov thaum sab hnub poob blot tso cai rau kev tshawb nrhiav thiab kom muaj nuj nqis ntawm cov protein tshwj xeeb los ntawm kev sib xyaw. Ob qho tib si muaj txiaj ntsig zoo hauv kev tshawb nrhiav cov protein.
SDS Page yog dab tsi?
SDS Nplooj ntawv yog cov txheej txheem gel electrophoresis siv rau kev sib cais cov protein. Nws yog feem ntau siv hauv biochemistry, noob caj noob ces, forensics thiab molecular biology. Thaum cov proteins tau muab rho tawm los ntawm cov qauv, lawv tau khiav ntawm gel ua los ntawm SDS thiab polyacrylamide. SDS yog cov tshuaj tua kab mob anionic uas yog siv los ua cov protein ntau (denature proteins) thiab muab cov nqi tsis zoo rau cov proteins uas sib npaug rau lawv cov molecular loj. Polyacrylamide ua cov khoom txhawb nqa rau cov gel. Denatured proteins uas tsis zoo them nqi tsiv mus rau qhov kawg ntawm lub cuab yeej los ntawm gel. Raws li qhov ntau thiab tsawg ntawm cov proteins, migrating speeds txawv ntawm cov proteins thiab kev sib cais tshwm sim. Yog li ntawd, SDS nplooj ntawv muaj txiaj ntsig zoo rau kev sib cais ntawm tus kheej cov proteins raws li lawv qhov ntau thiab tsawg.
Kev npaj ntawm polyacrylamide gel rau kev faib cov proteins yog ib kauj ruam tseem ceeb hauv SDS Page. Qhov tseeb concentration ntawm polyacrylamide thiab hom cross-linking tus neeg sawv cev siv tau muaj zog cuam tshuam rau lub cev lub cev ntawm cov gel, uas ua rau qhov sib cais ntawm cov protein sib txawv. Lub pore qhov ntau thiab tsawg ntawm cov gel yuav tsum tau tswj kom zoo rau kev sib cais zoo. Txawm li cas los xij, SDS Nplooj yog suav tias yog cov txheej txheem sib cais protein ntau.
SDS Page txheej txheem muaj kev txwv loj hauv kev txheeb xyuas cov protein. Txij li thaum SDS denatures proteins ua ntej kev sib cais, nws tsis tso cai rau kev tshawb pom ntawm enzymatic kev ua si, protein binding kev sib cuam tshuam, protein cofactors, thiab lwm yam.
Daim duab 01: SDS Page
Western Blot yog dab tsi?
Western blotting txheej txheem ua kom pom cov protein tshwj xeeb los ntawm cov protein sib xyaw thiab ntsuas qhov ntau thiab qhov hnyav ntawm cov protein. Western blot yog daim nyias nyias siv thaum lub sijhawm blotting kom tau txais daim iav duab ntawm cov qauv protein hauv SDS-polyacrylamide gel. Lub membrane uas siv rau sab hnub poob blotting feem ntau yog ua los ntawm nitrocellulose los yog polyvinylidene difluoride (PVDF). Lub membrane nrog cov protein hloov pauv tuaj yeem siv los txheeb xyuas cov protein uas xav tau. Nws yuav tsum muaj cov tshuaj tiv thaiv zoo rau kev tshawb nrhiav cov protein uas xav tau los ntawm hybridization. Cov tshuaj tiv thaiv kab mob khi rau nws cov tshuaj tiv thaiv tshwj xeeb thiab qhia pom muaj qhov xav tau antigen uas yog cov protein.
Kev xa cov proteins los ntawm SDS polyacrylamide gel mus rau sab hnub poob blot yog ua los ntawm electroblotting. Nws yog ib txoj hauv kev zoo thiab ceev ceev uas ua rau cov proteins electrophorese tawm ntawm cov gel thiab dhau mus rau nitrocellulose membrane (western blot).
Daim duab 02: Western Blot
Qhov txawv ntawm SDS Page thiab Western Blot yog dab tsi?
SDS Page vs Western Blot |
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| SDS Nplooj ntawv yog cov txheej txheem gel electrophoresis. | Western blot yog txheej txheem uas ua rau ntawm daim nyias nyias kom pom cov protein tshwj xeeb los ntawm kev sib xyaw. |
| SDS nplooj ntawv tso cai cais cov proteins raws li lawv qhov ntau thiab tsawg. | Western blot tso cai rau kev hloov pauv ntawm cov protein ntawm SDS nplooj gel yam tsis hloov nws cov qauv & tso cai rau kev sib xyaw nrog cov tshuaj tiv thaiv tshwj xeeb. |
| Disadvantages | |
| Protein denaturation, tus nqi siab, thiab muaj cov tshuaj neurotoxin yog qhov tsis zoo ntawm cov txheej txheem no. | Cov txheej txheem no siv sijhawm ntev thiab yuav tsum muaj kev paub txog tus kheej thiab cov kev tswj tshwj xeeb. |
Summary – SDS Page vs Western Blot
SDS nplooj ntawv thiab sab hnub poob blot yog ob txoj hauv kev koom nrog hauv kev txheeb xyuas cov protein. SDS Page tso cai yooj yim sib cais ntawm cov proteins ntawm ib gel raws li lawv qhov hnyav molecular. Western blot pab kom paub meej tias muaj thiab kom muaj nuj nqis ntawm cov protein tshwj xeeb los ntawm hybridization nrog cov tshuaj tiv thaiv tshwj xeeb. Qhov no yog qhov txawv ntawm SDS Page thiab western blot.
