cellproliferationassay

beforestart:

thawcellsfromliquidnitrogen,growin75ccflask(T75)inFischer"smediumMM(maintenancemedium)untilcontentgetscloudyandstartstoturnyellow(usuallywant~6x10⁶cellsperT75),assaywillbeperformedin24-well-plateatconcentrationof100,000cells/well,eachsampleintriplicate.

Onedaybeforeassay:

centrifugecells,bringbacktovolumewithFischer"sSM(stationarymedium).ItisimportanttoremoveallofMMfromcellssothatnegativecontrolislow.Ifwanted,cellscanberinsedinSMandspunagain.

Dayofassay:

spincells,bringtosmallervolumewithSMandcount,willwant100,000cells/0.5mL/well;allproteinsandcontrolsdoneintriplicate.Towellscontaining0.5mLcellsUSPensionadd:

• 0.5mLSM=negativecontrol
• 0.5mLMM=positivecontrol
• oPRL=positivecontrol(0.001ng-10ng(-100ng),intenfoldincrements,dilutedinSMto0.5mLtotalvolume),maxisreachedat1-10ng
• sampleproteinspreparedinthesameway(filtersterilizebeforeusing!)

incubateat37°Cfor72hrs,thenreadoncellcounter(SysmexMicrocellcounterCC110)

Cellcounter:

add400µLoftestsustanceto10mLHematall,countoncounter.ReADIngofcellcountx57,000iscellnumberin1mLoriginalcellculture

================  蚂蚁淘在线  ================

免责声明：本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容

版权声明：未经蚂蚁淘在线授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘在线”。违反上述声明者，本网将追究其相关法律责任。