Type
Recombinant
Description
TWEAKbelongstotheTNFfamilyofligands,andsignalsthroughTWEAKR,alsoknownasTNFRSF12A.TWEAKisexpressedinavarietyoftissues,includingtheadultheart,pancreas,skeletalmuscle,smallintestine,spleenandperipheralbloodlymphocytes.TWEAKhastheABIlitytoinduceNF-κBactivationandchemokinesecretion,andtoexertanapoptoticactivityincertaincells,suchasHT-29 humanadenocarcinomacellswhenculturedinthepresenceofIFN-γ.TWEAKalsopromotesproliferationandmigrationofendothelialcells.ThehumanTWEAKgeneencodesfora249 aminoacidtypeIItransmembraneprotein,whichcontainsa21 aminoacidcytoplasmicdomain,a21 aminoacidtransmembranedomain,anda207 aminoacidextracellulardomain.RecombinantHumanTWEAKisasoluble17.0 kDapolypeptide(154 aminoacidresidues)comprisingtheTNF-homologousregionofTWEAK,andisgeneratedbyproteolyticprocessingofthefulllengthmembrane-anchoredTWEAKprotein.
AminoAcidSequence
MKGRKTRARRAIAAHYEVHPRPGQDGAQAGVDGTVSGWEEARINSSSPLRYNRQIGEFIVTRAGLYYLYCQVHFDEGKAVYLKLDLLVDGVLALRCLEEFSATAASSLGPQLRLCQVSGLLALRPGSSLRIRTLPWAHLKAAPFLTYFGLFQVH
Source
E.coli
Purity
98%
BIOLOGicalActivity
Assay#1:TheED50asdeterminedbythedose-dependentstimulationofIL-8 productionbyHumanPBMCislessthan10 ng/ml. Assay#2:TWEAKweaklyinducesthedeathofHT29cellswhenculturedinthepresenceofIFN-ɣ.TheED50forthiseffectisbetween30–45 ng/ml.
Endotoxin
Endotoxinlevelis<0.1 ng/μgofprotein(<1EU/μg).
Reconstitution
Centrifugethevialpriortoopening.Reconstitutein10 mMSodiumPhosphate,pH7.5 toaconcentrationof1.0 mg/ml.Donotvortex.Forextendedstorage,itisrecommendedtofurtherdiluteinabuffercontainingacarrierprotein(example0.1%BSA)andstoreinworkingaliquotsat-20°Cto –80°C.
Storage/Expiration
–20°C
