Qhov Tseem Ceeb - Nitrocellulose vs PVDF
Western blotting yog ib txoj hauv kev uas tso cai rau kev tshawb nrhiav thiab ntsuas cov protein tshwj xeeb los ntawm cov qauv protein. Kev ntseeg tau ntawm cov txheej txheem yog nyob ntawm kev xaiv ntawm daim nyias nyias kom raug rau kev nqus cov protein los ntawm gel. Muaj ntau hom microporous membranes. Nitrocellulose thiab PVDF daim nyias nyias yog ob daim nyias nyias nyiam los ntawm cov kws tshawb fawb vim lawv cov cwj pwm tshwj xeeb tshaj lwm hom membranes. Kev xaiv ntawm nitrocellulose los yog PVDF kuj yog lwm qhov kev sib tw nyob rau sab hnub poob blotting. Ob leeg nitrocellulose thiab PVDF muaj peev xwm nqus tau cov protein ntau. Qhov sib txawv tseem ceeb ntawm nitrocellulose thiab PVDF daim nyias nyias yog tias nitrocellulose daim nyias nyias tsis muaj peev xwm tshem tawm cov tshuaj tiv thaiv kab mob thiab rov siv cov membrane rau cov tshuaj tiv thaiv kab mob thaum PVDF daim nyias nyias muaj peev xwm stripping thiab rov siv dua.
Nitrocellulose yog dab tsi?
Nitrocellulose yog ib lub polymer tsim los ntawm kev kho cellulose nrog nitric acid thiab yog siv los ua microporous daim nyias nyias nyob rau hauv molecular biology, tshwj xeeb tshaj yog rau blotting hom kev kawm xws li yav qab teb, qaum teb thiab sab hnub poob blotting. Qhov ntau thiab tsawg ntawm nitrocellulose daim nyias nyias muaj li ntawm 3 mus rau 20 µm. Nitrocellulose microporous microscous microscous full pom kev kuaj pom ntawm immunochemical cov tshuaj tiv thaiv tshwm sim rau saum npoo ntawm daim nyias nyias. Yog li ntawd, nitrocellulose daim nyias nyias yog nquag siv rau immobilization ntawm cov proteins thiab nrhiav kom tau cov proteins tshwj xeeb nyob rau sab hnub poob blotting. Nitrocellulose daim nyias nyias muaj peev xwm immobilize glycoproteins thiab nucleic acids.
Nitrocellulose daim nyias nyias yog nyiam nyob rau hauv lateral flow assays vim muaj ntau yam nta. Nitrocellulose membranes nqus cov proteins ntawm qhov siab. Cov kuab tshuaj uas siv los ntub cov membrane tsis txo cov nitrocellulose membrane nqus protein. Nitrocellulose daim nyias nyias tuaj yeem txiav tau yooj yim mus rau qhov xav tau gel loj thiab hloov cov protein los ntawm cov gel mus rau daim nyias nyias los ntawm hluav taws xob lossis capillary hloov. Nitrocellulose tso cai rau nrawm nrawm ntawm cov protein los ntawm daim nyias nyias uas muaj peev xwm khi tau siab. Nitrocellulose qhia txog kev txhim kho lub zog ntawm kev tuav. Lwm tus cwj pwm tshwj xeeb ntawm nitrocellulose daim nyias nyias yog tias nws tuaj yeem ua tau yooj yim glued nrog cov dej tsis muaj kuab tshuaj tiv thaiv adhesives mus rau ntau yam yas backings.
Daim duab 01: Nitrocellulose membrane rau western blotting
PVDF yog dab tsi?
Polyvinylidene difluoride (PVDF) yog fluoropolymer tsim los ntawm polymerization ntawm vinylidene difluoride thiab muaj peev xwm immobilization siab ntawm cov protein. Yog li ntawd, microporous daim nyias nyias ua los ntawm PVDF yog siv nyob rau hauv western blotting cov tswv yim los soj ntsuam cov proteins tshwj xeeb los ntawm cov sib tov ntawm cov proteins. PVDF daim nyias nyias kuj tseem siv tau rau kev tsom xam cov amino acid thiab protein sequencing. Cov yam ntxwv tseem ceeb tshaj plaws ntawm PVDF membrane tshaj nitrocellulose membrane yog tias nws tuaj yeem tau yooj yim tshem tawm cov tshuaj tiv thaiv thiab rov siv dua rau cov tshuaj tiv thaiv kab mob tom ntej.
PVDF daim nyias nyias yog tuab dua li nitrocellulose daim nyias nyias; li no, ntau resistant rau kev puas tsuaj thaum rov siv dua. PVDF membranes yog hydrophobic heev. Yog li ntawd, lawv yuav tsum tau tsau rau hauv methanol lossis isopropanol ua ntej siv.
Qhov txawv ntawm Nitrocellulose thiab PVDF yog dab tsi?
Nitrocellulose vs PVDF |
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| Nitrocellulose yog ib qho polymer tsim los ntawm cellulose. | PVDF yog fluoropolymer uas tsim los ntawm polymerization ntawm vinylidene difluoride. |
| membrane pore loj | |
| Qhov ntau qhov pore yog 0.1, 0.2 lossis 0.45μ | Qhov ntau qhov pore yog 0.1, 0.2 lossis 0.45μm |
| Protein Binding Peev xwm | |
| Nitrocellulose muaj peev xwm khi cov protein ntawm 80 txog 100 μg / cm 2. | PVDF muaj peev xwm khi cov protein ntawm 170 txog 200 μg / cm2. |
| xov xwm | |
| Qhov no muaj qhov rhiab heev piv rau PVDF. | Qhov no muaj qhov rhiab heev. |
| Tshawb nrhiav cov protein uas tsis tshua muaj txiaj ntsig | |
| Vim tias qhov rhiab heev ntawm cov khoom siv nitrocellulose tsawg, nws tsis tsim nyog rau kev kuaj pom cov proteins uas tsis tshua muaj. | Qhov no yog qhov tsim nyog rau kev kuaj pom cov proteins uas tsis tshua muaj vim nws qhov rhiab heev. |
| Background Noise | |
| No muaj suab nrov tom qab | Qhov no muaj suab nrov dua tom qab. |
| Kev Sib Tham nrog Proteins | |
| Protein molecules khi rau nitrocellulose daim nyias nyias los ntawm kev sib cuam tshuam hydrophobic. | Proteins khi rau PVDF daim nyias nyias los ntawm kev sib cuam tshuam hydrophobic thiab dipole. |
| Ntxim ntawm Membrane | |
| Nitrocellulose yog nkig thiab tawg. Txawm li cas los xij, cov qauv ntawm nitrocellulose muaj, thiab lawv tiv taus. | PVDF yog qhov ruaj khov dua thiab muaj tshuaj tiv thaiv ntau dua. |
| Muaj peev xwm rov qab siv dua | |
| Nitrocellulose tej zaum yuav muaj teeb meem hauv stripping thiab reprobing yam tsis poob lub teeb liab. | PVDF yog qhov zoo tagnrho rau kev rov hais dua thiab ua raws cov ntawv thov. |
| Kev tsim nyog | |
| Nitrocellulose yog qhov zoo tagnrho rau kev kuaj xyuas cov protein uas tsis muaj molecular. | PVDF yog qhov tsim nyog rau kev kuaj pom cov protein ntau dua molecular. |
| Lwm yam siv | |
| Nitrocellulose tuaj yeem siv rau kev tshuaj ntsuam nucleic acid thiab dot / slot blotting. | PVDF tuaj yeem siv rau kev sib cais ntawm cov protein thiab cov txheej txheem tshuaj ntsuam xyuas. |
| Tus nqi | |
| Qhov no yog pheej yig dua li PVDF membranes. | Qhov no kim dua li nitrocellulose membranes. |
| Yuav tsum tau ua ntej-wetting | |
| Nitrocellulose daim nyias nyias tsis tas yuav presoaking nrog methanol | PVDF daim nyias nyias yuav tsum tau presoaking nrog methanol. |
Summary – Nitrocellulose vs PVDF
Nitrocellulose daim nyias nyias yog thawj daim nyias nyias siv ua lag luam rau kev soj ntsuam xyuas sab nraud. Lawv muaj peev xwm nqus tau cov protein ntau. Yog li ntawd, nitrocellulose daim nyias nyias yog siv nyob rau hauv western blotting. PVDF yog lwm hom membrane siv nyob rau sab hnub poob blotting thiab nws kuj muaj peev xwm nqus tau cov protein ntau. Ob hom yog siv nyob rau sab hnub poob blotting rau kev tsom xam cov protein. Txawm li cas los xij, PVDF daim nyias nyias muaj cov yam ntxwv tshwj xeeb, ua rau lawv zoo dua li nitrocellulose membranes rau sab hnub poob blotting. Tab sis, nitrocellulose membranes yog qhov tsim nyog rau kev kuaj xyuas cov protein uas tsis muaj molecular hnyav, PVDF daim nyias nyias yog qhov tsim nyog rau kev kuaj pom cov protein ntau molecular. Qhov no yog qhov txawv ntawm nitrocellulose thiab PVDF membranes.
