Type
Recombinant
Description
CD40,amemberoftheTNFreceptorfamily,isacellsurfaceproteinexpressedonBcells,dendriticcells,monocytes,thymicepithelialcellsand,atlowlevels,onTcells.SignalingthoughCD40playsanimportantroleintheproliferationanddifferentiationofBcells,andiscriticalforimmunoglobulin(Ig)classswitching.Themembrane-anchoredCD40-LigandisexpressedalmostexclusivelyonactivatedCD4+Tlymphocytes.FailuretoexpressCD40Lleadsto“immunodeficiencywithhyper-IgM”,adiseasecharacterizedbyfailuretoproduceIgG,IgAandIgE.ThehumanCD40Lgenecodesfora261 aminoacidtypeIItransmembraneprotein,whichcontainsa22 aminoacidcytoplasmicdomain,a24 aminoacidtransmembranedomain,anda215 aminoacidextracellulardomain.ThesolubleformofCD40Lisan18 kDaproteincomprisingtheentireTNFhomologousregionofCD40LandisgeneratedinvivobyanintracellularproteolyticprocessingofthefulllengthCD40L.RecombinantHumansolubleCD40ligandisa16.3 kDaproteincontaining149 aminoacidresiduescomprisingthereceptorbindingTNF-likedomainof CD40L.
AminoAcidSequence
MQKGDQNPQIAAHVISEASSKTTSVLQWAEKGYYTMSNNLVTLENGKQLTVKRQGLYYIYAQVTFCSNREASSQAPFIASLWLKSPGRFERILLRAANTHSSAKPCGQQSIHLGGVFELQPGASVFVNVTDPSQVSHGTGFTSFGLLKL
Source
E.coli
Purity
98%
BiologicalActivity
Assay#1:DeterminedbythestimulationofIL-12 inductionbyhumanPBMCs.Assay#2:Determinedbythedose-dependentstimulationofIL-8 productionbyhumanPBMC. TheexpectedED50forthiseffectis5–10ng/ml.Note:ResultsmayvarywithPBMCdonors.
Endotoxin
Endotoxinlevelis<0.1 ng/μgofprotein(<1EU/μg).
Reconstitution
Centrifugethevialpriortoopening.Reconstitutein5–10mMSodiumPhosphate,pH7.5 toaconcentrationof0.1–1.0 mg/ml.Donotvortex.Forextendedstorage,itisrecommendedtofurtherdiluteinabuffercontainingacarrierprotein(example0.1%BSA)andstoreinworkingaliquotsat –20°Cto –80°C.
Storage/Expiration
–20°C