EntrezGeneSummary | TheproteinencodedbythisgeneisamemberoftheG-protein-coupledreceptorfamily.Thisproteinisareceptorforinterleukin8(IL8).ItbindstoIL8withhighaffinity,andtransducesthesignalthroughaG-proteinactivatedsecondmessengersystem.Knockoutstudiesinmicesuggestedthatthisproteininhibitsembryonicoligodendrocyteprecursormigrationindevelopingspinalcord.Thisgene,IL8RB,ageneencodinganotherhighaffinityIL8receptor,aswellasIL8RBP,apseudogeneofIL8RB,formageneclusterinaregionmappedtochromosome2q33-q36. |
IncubationConditions | RECOMMENDEDASSAYCONDITIONS:Membranesaremixedwithradioactiveligandandunlabeledcompetitor(seeFigures1and2forconcentrationstested)inbindingbufferinanonbinding96-wellplate,andincubatedfor1-2h.Priortofiltration,anFC96-wellharvestplate(Milliporecat.#MAHFC1H)iscoatedwith0.33%polyethyleneiminefor30min,thenwashedwith50mMHEPES,pH7.4,0.5%BSA.Bindingreactionistransferredtothefilterplate,andwashed3times(1mLperwellperwash)withWashBuffer.Theplateisdriedandcounted.
Bindingbuffer:50mMHepes,pH7.4,5mMMgCl2,1mMCaCl2,0.2%BSA,filteredandstoredat4°C
Radioligand:[125I]Interleukin-8(PerkinElmer#NEX277)
WashBuffer:50mMHepes,pH7.4,500mMNaCl,0.1%BSA,filteredandstoredat4°C.
Onevialcontainsenoughmembranesforatleast200assays(units),whereanunitistheamountofmembranethatwillyieldgreaterthan5-foldsignal:backgroundwith125I-labeledIL-8at0.1nM |