Kev tu thiab siv txoj kev ntawm ultrafiltration raj

Kev tu thiab siv txoj kev ntawm ultrafiltration raj

Lub luag haujlwm ntawm ultrafiltration tube:

1) Kev xav; 2) Kev xa tawm; 3) Hloov tsis tau; 4) Nws muaj cov nyhuv ntawm ib nrab purification.

1. Lub raj ultrafiltration tshiab tuaj yeem siv ncaj qha yam tsis muaj lwm yam kev kho mob;

2. Lub raj ultrafiltration tsis tuaj yeem ua kom tsis muaj menyuam ntawm qhov kub thiab txias;

3. Kev kho mob ntawm ultrafiltration raj thaum hloov cov concentrated protein

Lub raj ultrafiltration tuaj yeem rov siv dua. Soak nws hauv 0.1M NaOH rau 1-2h, ntxuav nws nrog dej, thiab tom qab ntawd ntxuav nrog cov tshuaj uas cuam tshuam (cov dej tsis muaj tshuaj lossis PBS) ntawm lub centrifuge rau 3 zaug, uas tuaj yeem siv los ua kom pom tseeb. tshiab protein;

4. Kev kho ntawm ib txwm muab tso rau ultrafiltration raj

Rau cov raj siv ultrafiltration uas nquag siv, tsau rau hauv 0.1M NaOH rau 1-2h, ntxuav nws nrog dej thiab tsau rau hauv dej kom tsis muaj menyuam. Nco ntsoov khaws cov lim membrane ntub thiab tiv thaiv kab mob kom tsis txhob loj hlob;

5. Kev kho ntawm ultrafiltration raj tso rau lub sijhawm ntev

Yog tias tsis siv lub raj ultrafiltration ntev, tsau rau hauv 0.1M NaOH rau 1-2h, ntxuav nws nrog dej, thiab tsau rau hauv 20 feem pua ​​​​ethanol cia. Nco ntsoov khaws cov lim membrane ntub thiab tiv thaiv kab mob kom tsis txhob loj hlob;

6. Centrifugal ceev siv rau ultrafiltration raj

Lub centrifugal ceev feem ntau yog 4000rpm (3000g feem ntau pom zoo, thiab qhov siab tshaj plaws tsis tuaj yeem tshaj 4000g). Tom qab centrifugation rau 5-8 feeb, 4ml qauv tuaj yeem ua kom muaj zog tsawg dua 1ml mus rau kaum tawm microliters; Yog hais tias qhov ceev yog siab dhau lawm, lub hom phiaj protein yuav poob, uas ua rau poob ntawm lub hom phiaj protein; Tsawg ceev, siv sijhawm ntev thiab ua haujlwm tsawg;

7. Kev ceev faj thaum rab phom taub hau txuas mus rau hauv lub raj ultrafiltration

Thaum ntxiv cov tshuaj protein los yog tsis muaj rau lub raj ultrafiltration, lub taub hau xiav rab phom tuaj yeem siv tau; Txawm li cas los xij, thaum lub hom phiaj tseem ceeb ntawm cov protein tau nqus los ntawm hauv qab ntawm lub raj ultrafiltration, lub taub hau daj yuav tsum tau siv; Thaum lub taub hau rab phom tau muab tso rau hauv lub raj ultrafiltration, nws yuav tsum tau tiv thaiv los ntawm kev kov lub lim membrane kom tsis txhob punctured lub lim membrane;

8. Txo cov protein adsorption poob ntawm ultrafiltration raj

Daim nyias nyias ntawm lub raj ultrafiltration tuaj yeem nqus cov protein, uas yuav txo tau cov hom phiaj protein li ntawm 30 feem pua; Txhawm rau kom tsis txhob muaj protein ntau, tom qab nqus cov kua dej ntawm lub hom phiaj, tshem tawm cov hauv qab ntawm lub raj ultrafiltration nrog cov protein tsis huv los ntxuav feem ntau ntawm cov protein adsorbed los ntawm lub lim membrane thiab txo qhov poob ntawm lub hom phiaj protein; Yog hais tias cov purified protein yog concentrated, xyuam xim hais tias lub concentrated ntim yuav tsum tsis txhob me heev; Feem ntau, tsis muaj teeb meem li cas centrifuged, txog 200 µ L protein tov yuav muab khaws cia rau hauv qab;

9. Yuav ua li cas los txhais qhov tsis ua hauj lwm ntawm ultrafiltration raj

Nyob rau hauv ib txwm siv, thaum lub lim membrane tsis punctured, cov concentrated protein tov tsis muaj protein ntau los yog tus nqi yog tsawg heev (xws li lub hom phiaj protein), thaum thawj lim dej muaj lub hom phiaj protein, nws qhia tau hais tias lub lim membrane yog tsis siv thiab. yuav tsum tau hloov nrog ib tug tshiab ultrafiltration raj.