Qhov txawv ntawm Coomassie thiab nyiaj staining yog tias coomassie staining yog cov txheej txheem protein staining uas siv coomassie ci ntsa iab xiav stain, thaum sliver staining yog cov txheej txheem protein staining uas siv cov xim nyiaj.
Kev sib cais thiab kev txheeb xyuas cov protein yog cov kauj ruam tseem ceeb hauv kev txheeb xyuas cov khoom noj. Lawv xav tau kev daws teeb meem ntawm cov protein ntau tom qab gel electrophoresis. Muaj ntau cov txheej txheem staining uas suav nrog cov stains sib txawv xws li anionic dyes (coomassie ci ntsa iab), hlau cations (imidazole-zinc), nyiaj stain thiab fluorescent dyes, thiab lwm yam. Qee zaum, cov xov tooj cua tuaj yeem siv tau. Kev xaiv ntawm cov txheej txheem staining nyob ntawm qhov yooj yim, muaj cov khoom siv duab hauv chav kuaj, thiab lwm yam.
Coomassie Staining yog dab tsi?
Coomassie staining yog cov txheej txheem protein staining uas siv coomassie ci ntsa iab xiav stain. Nws yog feem ntau hu ua coomassie xiav txheej txheem. Coomassie ci ntsa iab xiav yog qhov nrov tshaj plaws anionic protein-dye. Qhov no stain ib txwm stains yuav luag tag nrho cov proteins nrog cov khoom zoo linearity ntawm nruab nrab rhiab heev. Coomassie ci ntsa iab xiav stain khi tsis yog tshwj xeeb rau yuav luag txhua cov protein. Muaj ob qhov sib txawv coomassie ci ntsa iab xiav stain variants: R-250 thiab G-250. R-250 muaj lub sijhawm staining luv dua, thaum G-250 muaj nyob rau hauv ntau cov ntaub ntawv rhiab thiab zoo ib puag ncig.
Daim duab 01: Coomassie Staining
Coomassie dyes yog nrov heev hauv kev tshawb fawb loj spectrometry thiab kev tshawb fawb txog protein. Ntxiv mus, bio-safe coomassie stain yog ib qho kev tsim tsis muaj kev phom sij ntawm coomassie xiav G-250 uas tam sim no muaj nyob hauv khw. Qhov zoo ntawm cov qauv no yog nws tsuas yog xav tau dej rau yaug thiab destaining. Bio-safe coomassie stain muaj rhiab heev sib npaug rau cov pa coomassie xiav G-250 tab sis zoo dua li coomassie stain R-250. Tsis tas li ntawd, nws muaj cov txheej txheem yooj yim thiab nrawm dua li cov qauv Coomassie xiav G-250. Qhov tsis zoo ntawm coomassie xiav staining yog tias nws tsis tshua nkag siab piv rau nyiaj staining. Coomassie xiav staining yog kwv yees li 50-fold tsawg dua rhiab dua nyiaj staining. Txawm li cas los xij, vim nws qhov yooj yim hauv kev khi, nws nyiam nyob hauv ntau qhov kev tshawb fawb.
Nyiaj Staining yog dab tsi?
Silver staining yog cov txheej txheem staining protein uas siv cov xim nyiaj. Silver staining yog siv los stain ob qho tib si agarose thiab polyacrylamide gels. Cov nyiaj staining ntawm cov protein nyob rau hauv agarose gels yog thawj zaug tsim nyob rau hauv 1973 los ntawm Kerenyi thiab Galyas. Tom qab ntawd, nws tau hloov kho rau cov proteins hauv polyacrylamide gels uas siv hauv SDS-PAGE. Tam sim no, nyiaj staining kuj yog siv rau staining DNA lossis RNA. Yuav kom stains cov gels, cov gels yog incubated nrog silver nitrate tov nyob rau hauv txoj kev no. Silver staining stains cov chaw uas muaj protein ntau los ntawm xim av mus rau dub.
Daim duab 02: Nyiaj Staining
Qhov kev siv ntawm cov nyiaj staining nyob ntawm cov qauv ntawm cov protein. Tsis tas li ntawd, kev huv huv ntawm cov hlab ntsha siv thiab purity ntawm reagents tuaj yeem cuam tshuam qhov zoo ntawm cov nyiaj staining. Txawm li cas los xij, qhov tsis zoo ntawm cov nyiaj staining yog tias nws tsis tuaj yeem kuaj xyuas tag nrho cov proteins, tshwj xeeb tshaj yog glycoproteins thiab cov proteins nrog cov pab pawg hloov pauv loj txuas rau lawv cov saw hlau.
Dab tsi zoo sib xws ntawm Coomassie thiab Silver Staining?
- Coomassie thiab nyiaj staining yog ob txoj kev siv hauv cov protein staining.
- Lawv tuaj yeem ua rau cov proteins thiab DNA.
- Ob txoj kev staining muaj qhov tsis zoo.
- Cov txheej txheem no muaj cov kauj ruam zoo sib xws, xws li kho, staining thiab destaining.
Qhov txawv ntawm Coomassie thiab Silver Staining yog dab tsi?
Coomassie staining yog cov txheej txheem protein staining uas siv coomassie ci ntsa iab xiav, thaum sliver staining yog cov txheej txheem protein staining uas siv nyiaj stain. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm Coomassie thiab nyiaj staining. Tsis tas li ntawd, Coomassie staining tsis tshua muaj siab dua li nyiaj staining.
Cov hauv qab no infographic teev qhov sib txawv ntawm Coomassie thiab nyiaj staining nyob rau hauv daim ntawv tabular rau ib sab ntawm kev sib piv.
Summary – Coomassie vs Silver Staining
Txhawm rau pom cov protein, cov protein tshwj xeeb dye-binding lossis xim tsim cov tshuaj tiv thaiv tuaj yeem ua tau. Qhov no yog hu ua protein staining. Coomassie thiab nyiaj staining yog ob txoj kev siv hauv cov protein staining. Coomassie staining txheej txheem siv coomassie ci ntsa iab xiav stain thaum sliver staining siv nyiaj stain. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm Coomassie thiab nyiaj staining.
