Trypsinization yog cov txheej txheem ntawm kev sib cais ntawm tes siv trypsin, ib qho enzyme proteolytic uas zom cov proteins, kom tshem tawm cov hlwb los ntawm cov hlab ntsha uas lawv tab tom ua. Thaum ntxiv rau kab lis kev cai ntawm tes, trypsin rhuav tshem cov proteins uas ua rau lub hlwb ua raws li lub nkoj.
Vim li cas peb thiaj li sim cov hlwb?
Trypsinization feem ntau ua rau tso cai nkag ntawm cov hlwb mus rau lub thawv tshiab, kev soj ntsuam rau kev sim, lossis txo qis ntawm qhov sib npaug ntawm lub khob los ntawm kev tshem tawm ib feem pua. cov hlwb.
trypsin ua haujlwm li cas kom tshem tawm cov hlwb?
Trypsin / EDTA yog ib txoj hauv kev ua ke rau kev tshem tawm cov hlwb. Trypsin txiav cov adhesion proteins hauv cell-cell thiab cell-matrix kev sib cuam tshuam los ntawm kev txiav cov amino acid ntawm adhesion proteins tshwj xeeb ntawm lysine los yog aginine ntawm C-terminal yog hais tias upstream amino acid tsis proline.
Koj ua li cas thiaj tau trypsinized adherent cells?
txheej txheem
- Tshem tawm nruab nrab ntawm kab lis kev cai nkoj los ntawm kev thov thiab ntxuav lub monolayer nrog ntsev tsis muaj Ca2+ thiab Mg 2+ kom tshem tawm tag nrho cov kab mob ntshav qab zib. …
- Dispense txaus trypsin lossis trypsin / EDTA tov rau hauv kab lis kev cai hlab ntsha kom npog tag nrho cov monolayer ntawm cov hlwb thiab tso rau hauv 37 ° C incubator rau ~ 2 feeb.
Kuv yuav ua li cas lov tes taw trypsin hauv kab lis kev cai ntawm tes?
Ib zaug cov hlwb tshwm sim ntxiv 2 ntimntawm pre-warmed ua tiav kev loj hlob xov xwm kom inactivate trypsin. Maj mam disperse qhov nruab nrab los ntawm pippeting tshaj cell txheej nto ob peb zaug kom paub meej tias rov qab ntawm >95% ntawm cov hlwb. Rau cov kab lis kev cai dawb dawb, Soybean trypsin inhibitor (Cov Khoom No.
