Jackson/Peroxidase AffiniPure Rabbit AntiChicken IgY (IgG) (H+L)/1.5 ml/303035003

WholeIgGantibodiesareisolatedasintactmoleculesfromantiserabyimmunoaffinitychromatography.TheyhaveanFcportionandtwoantigenbindingFabportionsjoinedtogetherbydisulfidebondsandthereforetheyaredivalent.Theaveragemolecularweightisreportedtobeabout160kDa.ThewholeIgGformofantibodiesissuitableforthemajorityofimmunodetectionproceduresandisthemostcosteffective.

Basedonimmunoelectrophoresisand/orELISA,theantibodyreactswithwholemoleculechickenIgY.Italsoreactswiththelightchainsofotherchickenimmunoglobulins.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:None(Warning:Useofsodiumazideasapreservativewillsubstantiallyinhibittheenzymeactivityofhorseradishperoxidase.)

SuggestedWorkingConcentrationorDilutionRange:
ELISA:-1:5,000-1:100,000
WesternBlot:-1:5,000-1:100,000(non-ECL)/1:10,000-1:200,000(ECL)
Histo-/Cyto-Chemistry:-1:500-1:5,000

Dilutionfactorsarepresentedintheformofarangebecausetheoptimaldilutionisafunctionofmanyfactors,suchasantigendensity,permeability,etc.Theactualdilutionusedmustbedeterminedempirically.

HorseradishPeroxidase

Horseradishperoxidase(HRP)conjugatesarepreparedbyamodifiedNakaneandKawaoiprocedure(J.Histochem.Cytochem.1974.22,1084).Peroxidaseconjugatesarecommonlyusedforimmunohistochemistry,Westernblotting,andELISA.Affinity-purifiedanti-horseradishperoxidaseandconjugatesareavailablefordetectionofhorseradishperoxidaseantigenorforsignalamplificationofHRP-containingreagents.Forimmunostainingofmammaliancells,anadvantageofusinganti-horseradishperoxidaseisreducedbackground,sincetheantibodydoesnotrecognizetheendogenousperoxidase-likeenzymesfoundinthosecells.

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Jackson/Peroxidase AffiniPure Rabbit AntiChicken IgY (IgG) (H+L)/1.5 ml/303035003

HorseradishPeroxidase