Imikoreshereze nubwitonzi bwa ultrafiltration centrifuge tube
1) Hitamo umuyoboro ukwiye wa ultrafiltration.Menya ko UF membrane itandukanye murwego rwabo rwo kwihanganira imiti itandukanye.Mubisanzwe, umuyoboro wa ultrafiltration ufite 10 kDa ya molekuline yagabanijwe urashobora gutoranywa hamwe no kugabanya ibiro bya molekuline bitagomba kurenza 1/3 cyuburemere bwa poroteyine yinyungu, nka 35 kDa.Niba uburemere bwa molekuline ya poroteyine yinyungu bugera kuri 10 kd, umuyoboro wa ultrafiltration ufite uburemere bwa molekile uciwe na 3 KD urashobora gukoreshwa.
.Amazi noneho asukwa, ni ukuvuga amavuta ya poroteyine, hamwe nangahe yongeweho, ayo atarenze umurongo wera hejuru yigituba.Igikorwa kiroroshye, kandi umuyoboro wa ultrafiltration ugomba kubanziriza urubura mbere yo kongeramo proteine.
3) Byombi misa hamwe na centre de gravit yagombaga kugera kuringaniza.Menya umuvuduko wo kuzunguruka no kwihuta ntabwo byihuta cyane, bitabaye ibyo byangiza byimazeyo ultrafiltration membrane.Centrifugal ultrafiltration yatangiye (centrifuge yabanjirije dogere 4).Nyuma ya RPM ya centrifuges itandukanye yahinduwe g, byari bitandukanye.Kwihuta kwa centrifuge byahinduwe byibuze, bigabanya umuvuduko kuri membrane.Icyitonderwa, menya neza kuva muri centrifuge nyuma ya centrifuge igeze kumuvuduko wacyo, cyangwa niba ufite ikibazo, ntushobora kugikemura kunshuro yambere.Icyerekezo cya membrane kuri spindle cyahinduwe ukurikije amabwiriza (inguni ya centrifuge ni membrane to axis perpendicular).Mu mikoreshereze ifatika, umuvuduko rusange wo kuzenguruka uri munsi kurenza ayo mu mabwiriza, kugirango ubuzima bwumuyoboro wa centrifuge bushobora kwagurwa.
3) Byombi misa hamwe na centre de gravit yagombaga kugera kuringaniza.Menya umuvuduko wo kuzunguruka no kwihuta ntabwo byihuta cyane, bitabaye ibyo byangiza byimazeyo ultrafiltration membrane.Centrifugal ultrafiltration yatangiye (centrifuge yabanjirije dogere 4).Nyuma ya RPM ya centrifuges itandukanye yahinduwe g, byari bitandukanye.Kwihuta kwa centrifuge byahinduwe byibuze, bigabanya umuvuduko kuri membrane.Icyitonderwa, menya neza kuva muri centrifuge nyuma ya centrifuge igeze kumuvuduko wacyo, cyangwa niba ufite ikibazo, ntushobora kugikemura kunshuro yambere.Icyerekezo cya membrane kuri spindle cyahinduwe ukurikije amabwiriza (inguni ya centrifuge ni membrane to axis perpendicular).Mu mikoreshereze ifatika, umuvuduko rusange wo kuzenguruka uri munsi kurenza ayo mu mabwiriza, kugirango ubuzima bwumuyoboro wa centrifuge bushobora kwagurwa.
.Niba umuyoboro urekuye, ongera usuke urwego rwo hejuru hanyuma utembera mumiyoboro mishya kugirango utangire ultrafiltration.Kugirango umenye neza niba imiyoboro yabuze, centrifuge ya 10min hamwe na 5mgml ya BSA mbere yo gutembera, ikorera kuri kole ya proteine, cyangwa Bradford assay assay, hanyuma ukomeze wongereho proteine isigaye yibanze (ikora ku rubura kandi ikabuza poroteyine gushyuha) kugeza byose byibandwaho byongeweho.Witondere mugihe cya centrifugation niba haguye imvura ya poroteyine, biganisha ku gufunga imiyoboro.Niba imvura iguye, menya niba impamvu yihariye itera imvura ari proteine ikabije cyangwa buffer idakwiye;Iyambere irashobora gukemurwa na ultrafiltration yigituba cyinshi ultrafiltration icyarimwe, bikagabanya ubukana, hanyuma bigahinduka muguhindura ibisubizo bitandukanye kugeza igihe nta mvura ya proteine ibaye.
.Niba umuyoboro urekuye, ongera usuke urwego rwo hejuru hanyuma utembera mumiyoboro mishya kugirango utangire ultrafiltration.Kugirango umenye neza niba imiyoboro yabuze, centrifuge ya 10min hamwe na 5mgml ya BSA mbere yo gutembera, ikorera kuri kole ya proteine, cyangwa Bradford assay assay, hanyuma ukomeze wongereho proteine isigaye yibanze (ikora ku rubura kandi ikabuza poroteyine gushyuha) kugeza byose byibandwaho byongeweho.Witondere mugihe cya centrifugation niba haguye imvura ya poroteyine, biganisha ku gufunga imiyoboro.Niba imvura iguye, menya niba impamvu yihariye itera imvura ari proteine ikabije cyangwa buffer idakwiye;Iyambere irashobora gukemurwa na ultrafiltration yigituba cyinshi ultrafiltration icyarimwe, bikagabanya ubukana, hanyuma bigahinduka muguhindura ibisubizo bitandukanye kugeza igihe nta mvura ya proteine ibaye.
. inshuro zikurikiranye, hamwe nubunini bwanyuma bwanyuma bwanyuma bitewe nubushake bwa poroteyine wifuzaga, muri rusange ntabwo burenze 500ul, ariko no muri 200ul.Kugera kuri 1000 × cyangwa kurenga inshuro eshatu, cyane cyane impinduka za buffer, nkuko ubarwa byibuze 10 × cyangwa gutunganyiriza amajwi buri gihe.
. inshuro zikurikiranye, hamwe nubunini bwanyuma bwanyuma bwanyuma bitewe nubushake bwa poroteyine wifuzaga, muri rusange ntabwo burenze 500ul, ariko no muri 200ul.Kugera kuri 1000 × cyangwa kurenga inshuro eshatu, cyane cyane impinduka za buffer, nkuko ubarwa byibuze 10 × cyangwa gutunganyiriza amajwi buri gihe.
Igihe cyo kohereza: Ugushyingo-09-2022