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AAT Bioquest/Cell Navigator™ F-Actin Labeling Kit *Orange Fluorescence*/22663/500 Tests
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 546/575 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellBIOLOGy LabelingCells |
| Related | FluorescenceImaging BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1.Prepare1XiFluor™546-Phalloidinworkingsolution:
Add10µLofiFluor™546-Phalloidin(ComponentA)to10mLofLabelingBuffer(ComponentB).
Note1:TheunusediFluor™546-Phalloidinstocksolution(ComponentA)shouldbealiquotedandstoredat-20ºC.Protectfromlight.
Note2:Differentcelltypesmightbestaineddifferently.TheconcentrationofiFluor™546-Phalloidinworkingsolutionshouldbepreparedaccordingly.
2.Stainthecells:
2.1 Performformaldehydefixation.Incubatethecellswith3.0–4.0%formaldehydeinPBSatroomtemperaturefor10–30minutes.
Note:Avoidanymethanolcontainingfixativessincemethanolcandisruptactinduringthefixationprocess.Thepreferredfixativeismethanol-freeformaldehyde.
2.2 Rinsethefixedcells2–3timesinPBS.
2.3 Optional:Add0.1%TritonX-100inPBSintofixedcells(fromStep2.2)for3to5minutestoincreasepermeability.Rinsethecells2–3timesinPBS.
2.4 Add100μL/well(96-wellplate)ofiFluor™546-Phalloidinworkingsolution(fromStep1)intothefixedcells(fromStep2.2or2.3),andstainthecellsatroomtemperaturefor15to60minutes.
2.5 RinsecellsgentlywithPBS2to3timestoremoveexcessdyebeforeplatesealingandimagingbyusingTRITCchannel.
| References&Citations |  CitationExplorer |
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