Ib zaug cov hlwb tshwm sim ntxiv 2 ntim ntawm pre-warmed ua tiav kev loj hlob xov xwm kom tsis ua haujlwm trypsin. Maj mam disperse qhov nruab nrab los ntawm pippeting hla lub cell txheej nto ob peb zaug kom paub meej tias rov qab ntawm >95% ntawm cov hlwb.
Kuv yuav ua li cas thiaj tsis ua haujlwm trypsin?
Lub trypsin digestion tuaj yeem nres los ntawm freezing los yog txo qis pH ntawm cov tshuaj tiv thaiv hauv qab pH 4 los ntawm kev ntxiv formic, acetic, los yog trifluoroacetic acid (trypsin yuav rov ua haujlwm thaum lub pH nce siab tshaj pH 4). Cov qauv digested tuaj yeem khaws cia ntawm -20 ° C.
Dab tsi neutralizes trypsin?
A tsis muaj calcium thiab magnesium ions yuav ua rau trypsin qeeb. FBS yog txawm li cas los xij txoj hauv kev zoo dua los tiv thaiv kev ua haujlwm trypsin.
Puas cov ntshav tsis ua haujlwm trypsin?
Serum inactivates residual trypsin seemlos ntawm enzymatic digestion ntawm ob lub raum thiab cov proteolytic enzymes tom qab synthesized los ntawm cov hlwb. Freshly trypsinized hlwb tuaj yeem loj hlob mus rau monolayers thaum tsis muaj ntshav qab zib tau muab tias lawv tau rov qab ntxuav kom tshem tawm cov seem seem ntawm trypsin.
Nws siv ntau npaum li cas xov xwm los txiav txim siab trypsin?
Tsuav koj siv tsawg kawg yog 1: 1 piv ntawm 5-10% cov ntshav -muaj nruab nrab nrog koj cov trypsin yuav tsum muaj ntau tshaj qhov txaus inhibition, ua raws li centrifugation thiab nruab nrab pauv nrog cov nruab nrab tshiab.
