Description:
CLIP-Cell™505isagreenfluorescentsubstratethatcanbeusedtolabelCLIP-tag™fusionproteinsinsidelivingcells,oncellsurfacesorinvitro.Thiscell-permeablesubstrate(BC-505)isbasedontheDyomicsdyeDY-505andissuitableforstandardfluoresceinfiltersets.Ithasanexcitationmaximumat504nmandemissionmaximumat532nm.Thispackageincludes50nmolofCLIP-Cell505substrate,sufficienttomake10mlofa5µMCLIP-tagfusionproteinlabelingsolution.
TheCLIP-tagproteinlabelingsystemenablesthespecific,covalentattachmentofvirtuallyanymoleculetoaproteinofinterest.CLIP-tagisaproteintagbasedonhumanO6-alkylguanine-DNAalkyltransferase(hAGT).CLIP-tagsubstratesarederivativesofbenzylcytosine(BC).Inthelabelingreaction,thesubstitutedbenzylgroupofthesubstrateiscovalentlyattachedtothereactivecysteineofCLIP-tagformingastablethioetherbond. AlthoughCLIP-tagisbasedonthesameproteinasSNAP-tag®,thebenzylcytosinesubstratesformaseparateclassofsubstrates,differentfromthebenzylguaninesubstratesrecognizedbySNAP-tag.CLIP-tagandSNAP-tagcanbeusedfororthogonalandcomplementarylabelingoftwoproteinssimultaneouslyinthesamecells.
Therearetwostepstousingthissystem:subcloningandexpressionoftheproteinofinterestasaCLIP-tagfusionwiththeCLIP-tagfusionproteinsisdescribedinthedocumentationsuppliedwithCLIP-tagplasmids.Thelabelingofthefusionproteinsisdescribedinthisdocument.
Notes:
Storage:CLIP-Cell505shouldbestoredat-20°C(longterm)orat4°Cinthedark(shortterm,lessthan4weeks).Protectthesubstratefromlightandmoisture.Withproperstorageat-20°Cthesubstrateshouldbestableforatleastthreeyearsdryor3monthsdissolvedinDMSO.