WholeIgGantibodiesareisolatedasintactmoleculesfromantiserabyimmunoaffinitychromatography.TheyhaveanFcportionandtwoantigenbindingFabportionsjoinedtogetherbydisulfidebondsandthereforetheyaredivalent.Theaveragemolecularweightisreportedtobeabout160kDa.ThewholeIgGformofantibodiesissuitableforthemajorityofimmunodetectionproceduresandisthemostcosteffective.

Basedonimmunoelectrophoresisand/orELISA,theantibodyreactswithwholemoleculeratIgG.Italsoreactswiththelightchainsofotherratimmunoglobulins.Noantibodywasdetectedagainstnon-immunoglobulinserumproteins.Theantibodymaycross-reactwithimmunoglobulinsfromotherspecies.
PhysicalState:Freeze-driedsolid
Storage:

Storefreeze-driedpowderat2-8°C.Whenreadytouse,rehydratewithindicatedvolumeofd.waterandcentrifugeifnotclear.Productisstableforabout6weeksat2-8°Casanundilutedliquid.Prepareworkingdilutionfresheachday.Forextendedstorageafterrehydration,addanequalvolumeofglycerol(ACSgradeorbetter)forafinalconcentrationof50%,andstoreat-20°Casaliquid.Note:aftertheadditionofglycerol,theconcentrationofproteinandbuffersaltsisone-halfoftheoriginal.Alternatively,aliquotandfreezetheproductat-70°Corbelowintheabsenceofglycerol.Avoidrepeatedfreezingandthawing.
Expirationdate:oneyearfromdateofrehydration.However,theexpirationdatemaybeextendediftheproductisstoredaccordingtotherecommendationandthetestresultsareacceptableforitsintendeduse.


Purity:Theantibodywaspurifiedfromantiserabyimmunoaffinitychromatographyusingantigenscoupledtoagarosebeads.
Buffer:0.01MSodiumPhosphate,0.25MNaCl,pH7.6
Preservative:0.05%SodiumAzide

SuggestedWorkingConcentrationorDilutionRange:
Histo-/Cyto-Chemistry:-1:100-1:800
FlowCytometry:-1:100-1:800

Dilutionfactorsarepresentedintheformofarangebecausetheoptimaldilutionisafunctionofmanyfactors,suchasantigendensity,permeability,etc.Theactualdilutionusedmustbedeterminedempirically.

CyanineCy™3

Amax:550Emax:570nm

Cy3isbrighter,morephotostable,andgiveslessbackgroundthanotherorange-redfluorescingdyeconjugates.Cy3conjugatescanbeexcitedmaximallyat550nm,withpeakemissionat570nm.Forfluorescencemicroscopy,Cy3canbevisualizedwithtraditionaltetramethylrhodamine(TRITC)filtersets,sincetheexcitationandemissionspectraarenearlyidenticaltothoseofTRITC.WerecommendCy3asabrighteralternativetoTRITC.Cy3canbeexcitedtoabout50%ofmaximumwithanargonlaser(514nmor528nmlines),ortoabout75%ofmaximumwithahelium/neonlaser(543nmline)ormercurylamp(546nmline).Cy3hasbeenusedwithfluoresceinfordoublelabeling;however,theuseofanarrowband-passemissionfilterforfluoresceinisrecommendedtominimizeCy3fluorescenceintheFITCfilterset.Cy3canalsobepairedwithAlexaFluor®647formultiplelabelingwhenusingaconfocalmicroscope.However,abetterchoiceformultiplelabelingisRhodamineRed-Xbecauseitsfluorescenceismidwaybetweenagreenfluorescingdye(likeAlexaFluor®488)andafar-red-fluorescingdyelikeAlexaFluor®647.