Qhov Tseem Ceeb Qhov txawv - Stacking Gel vs Separating Gel
Cov ntsiab lus stacking gel thiab cais gel yog siv los piav qhia txog cov txheej txheem SDS-PAGE. SDS-PAGE los yog sodium dodecyl sulfate-polyacrylamide gel electrophoresis yog cov txheej txheem kuaj uas siv los cais cov protein molecules raws li lawv qhov hnyav molecular. Txoj kev xav tom qab cov txheej txheem yog cov proteins uas muaj qhov sib txawv molecular hnyav qhia qhov sib txawv ntawm kev tsiv teb tsaws; Tsawg molecular hnyav molecules migrate sai dua whereas siab molecular hnyav molecules migrate maj mam. Qhov nruab nrab rau kev tsiv teb tsaws yog gel. Muaj ob hom gels hu ua stacking gel thiab cais gel. Qhov sib txawv tseem ceeb ntawm stacking gel thiab cais gel yog tias pH ntawm stacking gel yog 6.8 whereas pH ntawm cov gel cais yog 8.8.
Stacking Gel yog dab tsi?
Stacking gel yog ib qho tsis tshua muaj siab polyacrylamide gel uas tau muab tso rau sab saum toj ntawm ntau cov tshuaj daws teeb meem (sib cais gel) hauv SDS-PAGE txheej txheem. Lub stacking gel yog siv los txhim kho qhov kev daws teeb meem ntawm electrophoresis. Qhov kev daws teeb meem nce ntxiv vim qhov sib txawv ntawm qhov sib txawv ntawm cov gel stacking thiab daws cov gel uas cuam tshuam rau cov proteins hauv cov qauv. Txij li thaum lub concentration ntawm polyacrylamide nyob rau hauv stacking gel yog tsawg, lub pore loj dua. Qhov no kuj pab txhawb kev sib cais.
Lub stacking gel yog xoob polyacrylamide gel nrog cov pores loj txog li 7% polyacrylamide. Cov pores no tsis ua raws li qhov cuam tshuam loj rau cov protein molecules loj. Yog li no gel cuam tshuam me ntsis ntawm kev txav mus los ntawm cov proteins. Qhov no ua rau kev sib cais raws li kev txav mus los thiab qhov loj ntawm cov protein los ntawm kev tsom lawv ntawm ob gels.
Daim duab 01: SDS-PAGE Acrylamide Gel
Tus pH ntawm cov gel stacking yog 6.8. Nws pH yog acidic dua li ntawm kev daws gel los ntawm 2 pH units. Qhov pH no qhia txog lub zog ionic qis dua li qhov hluav taws xob ntau dua. Qhov no ua rau muaj kev txav mus los ntawm cov protein ntau dua li lwm cov khoom siv hluav taws xob muaj nyob hauv gel.
Dab tsi yog Separating Gel?
Sib cais cov gel lossis daws cov gel ntawm SDS-PAGETechnique yog cov tshuaj muaj zog polyacrylamide gel uas tau muab tso rau saum cov gel uas tsis tshua muaj siab. Cov gel sib cais muaj ntau ntawm polyacrylamide (10%). Lub pH ntawm cov gel no yog khaws cia raws li pH=8.8, uas yog qib pH siab dua li cov gel stacking.
Daim duab 02: Ib Gel Electrophoresis Apparatus
Thaum cov protein molecules ncav cuag cov gel sib cais, kev tsiv teb tsaws ntawm cov molecules yog qeeb vim tias cov gel cais tawm yog cov gel siab concentration nrog me me pore loj uas tuaj yeem ua qhov cuam tshuam loj rau kev txav ntawm cov protein molecules.. Qhov no qeeb qeeb tso cai rau lwm cov proteins tau tsiv mus qeeb kom ncav cuag, ua rau muaj qhov nqaim, muaj zog hauv nruab nrab ntawm ob lub gels.
Kev sib txheeb ntawm Stacking Gel thiab Separating Gel yog dab tsi?
Ob qho tib si Stacking Gel thiab Separating Gel hom yog siv rau hauv tib lub cuab yeej
Qhov txawv ntawm Stacking Gel thiab Separating Gel yog dab tsi?
YStacking Gel vs Separating Gel |
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| Stacking gel yog ib qho tsis tshua muaj siab polyacrylamide gel uas tau muab tso rau saum cov tshuaj daws teeb meem ntau dua (sib cais gel) hauv SDS-PAGE txheej txheem. | Sib cais gel lossis daws cov gel ntawm SDS-PAGE cov txheej txheem yog cov tshuaj muaj zog polyacrylamide gel uas tau muab tso rau saum cov gel uas tsis tshua muaj siab. |
| Chaw | |
| Stacking gel yog muab tso rau ntawm kev daws teeb meem (sib cais) gel. | Sib cais gel muab tso rau hauv qab ntawm lub thawv siv rau gel electrophoresis. |
| Qhov concentration ntawm stacking gel yog siab. | Qhov concentration ntawm kev sib cais gel yog qis. |
| Polyacrylamide Cov ntsiab lus | |
| Stacking gel muaj txog 7% polyacrylamide. | Sib cais gel muaj txog 10% polyacrylamide. |
| pH | |
| PH ntawm cov gel stacking yog 6.8. | Tus pH ntawm cov gel cais yog 8.8. |
| Pore Size | |
| qhov pore loj muaj nyob rau hauv stacking gel. | me me pore loj muaj nyob rau hauv cais gel. |
| Resolution | |
| Stacking gel muab daws teeb meem zoo dua. | Sib cais gel muab daws teeb meem tsis zoo. |
Summary – Stacking Gel vs Separating Gel
Stacking gel thiab cais gel yog ob hom polyacrylamide gels siv kom tau txais kev sib cais zoo ntawm cov protein molecules hauv cov qauv muab. Qhov txawv ntawm stacking gel thiab cais gel yog tias pH ntawm stacking gel yog 6.8 whereas pH ntawm cov gel cais yog 8.8.
