| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | None/None |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | |
| Category | MicroBIOLOGy FluorescenceImaging |
| Related | BiochemicalAssays |
1.ProteinPrecipitation:
1.1 Prepareaclearsolutionsamplewithproteinconcentrationlessthan50mg/mLafterhomogenizationandcentrifugation.Keepproteinsamplesonice.
Note:Highproteinconcentrationsamplesmightneedtobediluted.
1.2 Add10μLofcoldTCA(ComponentA)into100μLofproteinsampleandvortexbrieflytomixwell.
1.3 Keepthesampleonicefor10minutes.Centrifugeat12,000rpmfor5minutes.Transfersupernatant(deproteinizedsample)toanothertube.
Note:ItisrecommendedtoneutralizesamplesinTCA(Step2.1)anduseforrelevantassaysimmediately.However,deproteinizedsamplesinTCAmaybestoredat-70ºCforuptoonemonth.
2.TCANeutralization:
2.1 ToneutralizeexcessTCA,add10μLofcoldNeutralizationSolution(ComponentB)intothecollectedsupernatantandvortexbrieflytomixwell.
2.2 Keepthesamplesonicefor5minutes.Samplesarenowdeproteinizedandneutralized.
2.3 (Optional)Readabsorbanceat280nm(OD)tocheckifthesamplesaredeproteinized.
| References&Citations |  PrinterFriendlyVersion |
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