1. Lub hauv paus ntsiab lus ua haujlwm ntawm G418 kuaj 293T hlwb
G418 yog ib qho tshuaj tua kab mob neomycin uas cuam tshuam nrog kev sib txuas ntawm cov protein nyob hauv lub cev, yog li ua rau cov cell tuag. Txawm li cas los xij, cov hlwb nrog cov noob neomycin tsis kam tuaj yeem tsim cov protein tiv thaiv los tiv thaiv cov tshuaj lom ntawm G418. Yog li ntawd, nyob rau hauv kab lis kev cai nruab nrab uas muaj G418, tsuas yog cov hlwb uas muaj cov noob neomycin tsis kam tuaj yeem muaj sia nyob thiab loj hlob.
2. Cov nyhuv ntawm G418 concentration ntawm 293T hlwb
Cov nyhuv ntawm G418 concentration ntawm 293T hlwb yog multifaceted, nrog rau kev loj hlob ntawm tes, cell viability, cell morphology thiab cell resistance. Cov teebmeem tshwj xeeb ntawm G418 concentration ntawm 293T hlwb feem ntau tshwm sim hauv cov hauv qab no:
1. Kev loj hlob ntawm tes: Nyob rau hauv qis G418 concentrations (xws li 0.25 mg/ml), 293T hlwb tuaj yeem loj hlob ib txwm, cov cell ceev yuav maj mam nce, thiab cov morphology tseem zoo. Nrog rau qhov nce ntawm G418 concentration (xws li 0.5 mg/ml), qhov kev loj hlob ntawm tes pib qeeb thiab cov cell ceev nce qeeb. Thaum G418 concentration nce mus txog 1 mg / ml, qee lub hlwb pib tuag thiab cell ceev txo qis. Txuas ntxiv mus nce G418 concentration (xws li 2 mg / ml), kev tuag ntawm tes yog qhov hnyav dua thiab cov xov tooj ntawm tes txo qis. 2. Cell viability: Qhov siab dua G418 concentration, lub cell viability qis dua. Ntawm qhov tsis tshua muaj siab ntawm G418 (xws li 0.25 mg / ml), cell viability siab, cell morphology yog tag nrho, thiab staining yog uniform. Raws li qhov concentration ntawm G418 nce, cell viability maj mam txo, cell morphology ua tsis xwm yeem, thiab staining tsis sib xws. Ntawm qhov siab ntawm G418 (xws li 2 mg / ml), cell viability yog tsawg, cell morphology raug mob hnyav, thiab staining yog txawv txav.
3. Cell morphology: Qhov siab dua qhov concentration ntawm G418, qhov tsis xwm yeem ntawm cov cell morphology ntau dua. Ntawm qhov tsis tshua muaj siab ntawm G418, lub cell morphology yog plump, spindle-shaped los yog tiaj tus. Raws li qhov concentration ntawm G418 nce, lub cell morphology ua tsis xwm yeem, lub xov tooj ntawm tes poob qis, thiab cov npoo ntawm tes tsis meej. Ntawm qhov siab ntawm G418, lub cell morphology raug puas tsuaj loj, lub cev ntawm tes me me, thiab cov npoo ntawm tes tau tawg.
4. Cell tsis kam: qhov siab dua ntawm G418, qhov muaj zog ntawm tes tsis kam. Ntawm qhov tsis tshua muaj siab ntawm G418, cov hlwb muaj rhiab heev rau G418 thiab ua rau cov cell tuag. Raws li qhov concentration ntawm G418 nce, cov hlwb maj mam txhim kho kev tiv thaiv thiab cell ciaj sia taus nce. Ntawm qhov siab ntawm G418, kev tiv thaiv ntawm tes mus txog nws lub ncov, thiab cov hlwb muaj peev xwm tiv taus cov toxicity ntawm siab concentrations ntawm G418.
Yog li, rau 293T hlwb, qhov kev tshuaj ntsuam xyuas ntawm G418 feem ntau yog nruab nrab ntawm 1-2 mg / ml. Qhov kev xav tau no tuaj yeem hloov kho raws li kev sim xav tau. Feem ntau hais lus, qhov siab dua qhov kev tshuaj ntsuam xyuas, qhov kev tshuaj ntsuam zoo dua, tab sis nws kuj yuav ua rau qee yam kev puas tsuaj rau lub hlwb. Yog li ntawd, thaum xaiv qhov kev ntsuam xyuas concentration, nws yog ib qho tsim nyog los ntsuas qhov kev sim lub hom phiaj thiab kev ua siab ntev ntawm tes. Hauv kev ua haujlwm tiag tiag, koj tuaj yeem pib nrog qhov qis dua, xws li 0.5 mg / ml, saib xyuas kev loj hlob ntawm tes, thiab maj mam nce ntxiv kom txog rau thaum koj pom qhov kev ntsuam xyuas tsim nyog.