Product name :Cell Counting Kit (CCK)

Product Code:K009

CCK is a one-bottle solution;no premixing of components is required.CCK ,being nonradioactive, allows sensitive

colorimetric assays for the determination of the number of viable cells in cell proliferation and cytotoxicityassays.

WST-8 is reduced by dehydrogenases in cells to give an orange colored product(formazan),which is soluble in

the tissue culture medium(figure2).The amount of the formazan dye generated by dehyhrogenases in cells isdirectly proportional to the number of living cells.

Figure3. Different of CCK,XTT,MTS and MTT

Figure3showsthatthecellproliferation assay usingCCK correlateswell withthe[3H]-thymidineincorporationassay.

Thus,theCCKassaycanalsobesubstitutedforthe[3H]-thymidineincorporationassay.Asishigherthanthatusing

othertetrazoliumsaltssuchasMTT,XTT,MTSorWST-1.

50tests:0.5ml×1tube

100tests: 1ml×1tube

500tests: 5ml×1bottle

1000tests:10ml×1bottle

3000tests:30ml×1bottle

CCKisstableoveroneyearat4℃withprotectionfromlight.Storeitat-20℃forlongerstorage.Repeatedthawing

andfreezingcausesanincreaseinthebackground,which interferes with theassay.Please store thekitat4℃for

frequentuse.

-platereader(450nmfilter)

-96-wellplate

-CO2incubator

-10uland100-200ulmulti-channelpipettes

NumberofCellsDetermination

1.Inoculatecellsuspension(100ul/well）ina96-wellplate.Pre-incubatetheplateinahumidifiedincubator(e.g.,at37℃，

5%CO2).

2.Add10uloftheCCKsolutiontoeachwelloftheplate.

Becarefulnottointroducebubblestothewells,sincetheyinterferewiththeO.D.Reading.

4.Incubatetheplatefor1-4hoursintheincubator.

5.Measuretheabsorbanceat450nmusingamicroplatereader.

Tomeasuretheabsorbancelater,add10μlof1%w/vSDSor0.1MHCltoeachwell,covertheplateandstore itwith

protectionfromlightatroomtemperature.No absorbance change should beobservedfor24hours.

CellProliferationandCytotoxicityAssay

1.Dispense100μlofcellsuspension(5000cells/well)ina96-wellplate.Pre-incubatetheplatefor24hoursina humid

-ifiedincubator(e.g.,at37°C,5%CO2).

2.Add10μlofvariousconcentrationsofsubstancestobetestedtotheplate.

3.Incubatetheplateforanappropriatelengthoftime(e.g.,6,12,24or48hours)intheincubator.

4.Add10μlofCCKsolutiontoeachwelloftheplate.

Becarefulnottointroducebubblestothewells,sincetheyinterferewiththeO.D.reading.

5.Incubatetheplatefor1-4hoursintheincubator.

6.Measuretheabsorbanceat450nmusingamicroplatereader.

Tomeasuretheabsorbancelater,add10μlof1%w/vSDSor0.1MHCltoeachwell,covertheplateandstoreitwith

protectionfromlightatroomtemperature.Noabsorbancechangeshouldbeobservedfor24hours.

Cell Counting Kit(CCK) allows very convenient assays by utilizing ZETA LIFE‘s highlywater-soluble tetrazolium salt.