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Prospec/Leptin-B Tilapia/Leptin-B Tilapia Recombinant/20µg/CYT-1110-20µg-20µg
Cataloguenumber
CYT-1110
Introduction
Leptinisaproteinhormone.ItismainlyproducedinADIposecellsthatregulateenergyhomeostasisbyrestraininghunger.Leptintiestonuclearreceptorsinthehypothalamus(arcuatenucleus).SimilartoinsulinresistanceintypeIIdiabetes,inobesitythereisadecreaseinthesensitivitytowardsleptin,endinginafailuretoidentifysatiety,eveninhighlevelsofenergystoresorleptinitself.Full-lengthCDNAencoding2leptinsequences(tLepAandtLepB)and1leptinreceptorsequence(tLepR)existsintilapia(Oreochromisniloticus).Thefull-lengthcDNAoftLepRis3423bp,encodingaproteinof1140aminoacidwhichcontainedallfunctionallyimportantdomainsconservedamongvertebrateleptinreceptors.ThecDNAsoftLepAandtLepBare486bpand459bpinlength,encodingproteinsof161aaand152aa,respectively.Thethree-dimensionalstructuresoftLepAandtLepBdemonstratesstrongconservationoftertiarystructurewiththatofhumanleptincomprisedof4helixes.
Description
Leptin-BTilapiaRecombinantproducedinE.Coliisasingle,non-glycosylatedpolypeptidechaincontaining152aminoacidsandhavingamolecularmassof15,243Dalton.
TheLeptin-BTilapiaispurifiedbyproprietarychromatographictechniques.
Source
PhysicalAppearance
Formulation
Lyophilizedfromaconcentrated(1mg/ml)solutioncontainingNaHCO3at1:2salt:proteinratio.
Solubility
ItisrecommendedtoreconstitutethelyophilizedLeptin-BTilapiainsterilewateror0.4%NaHCO3adjustedtopH8-9,notlessthan100µg/ml,whichcanthenbefurtherdilutedtootheraqueoussolutions.
StABIlity
LyophilizedLeptin-BTilapiaalthoughstableatroomtemperaturefor3weeks,shouldbestoreddesiccatedbelow-18°C.UponreconstitutionLeptin-BTilapiashouldbestoredat4°Cbetween2-7daysandforfutureusebelow-18°C.Forlongtermstorageitisrecommendedtoaddacarrierprotein(0.1%HSAorBSA).Pleasepreventfreeze-thawcycles.
Purity
Greaterthan95.0%asdeterminedby:(a)Gelfiltrationanalysis.(b)AnalysisbySDS-PAGE.
Aminoacidsequence
ThefirstsixN-terminalaminoacidsofrecombinantTilapialeptinBareAla-Leu-Leu-Thr-Lys-Gly.
BIOLOGicalActivity
TilapialeptinswerefoundtobebiologicallyactiveinpromotingproliferationofBAF/3cellsstablytransfectedwiththelongformofhumanleptinreceptor,buttheiractivitywaslowerthanthatofmammalianleptin.FurThermore,theTilapialeptinswerebiologicallyactiveinpromotingSTAT‐LUCactivationinCOS7cellstransfectedwithTilapialeptinreceptorbutnotincellstransfectedwithhumanleptinreceptor.TilapiaLeptinAwasmoreactivethanTilapiaLeptinB.
Proteincontent
ProteinquantitationwascarriedoutbyUVspectroscopyat280nmusingtheabsorbencyvalueof0.19for1mg/mlLeptin-BTilapiaastheextinctioncoefficientfora0.1%(1mg/ml)solutionatpH8.0.ThisvalueiscalculatedbytheDNAmancomputeranalysisprogramofproteinsequences.
SafetyDataSheet
SDS
