Overview:ProductNameHSP25/HSP27AntibodyDescriptionMouseAnti-HumanHSP25/HSP27MonoclonalIgG1KappaSpeciesReactivityDog,Human,Mouse,Rat,Bovine,GuineaPig(Caviaporcellus),Hamster,SheepApplicationsWB,IHC,ICC/IF,IP,FCM,FACSAntibodyDilutionWB(1:5000),ICC/IF(1:200),IHC(1:100);optimaldilutionsforassaysshouldbedeterminedbytheuser.HostSpeciesMouseImmunogenSpeciesHumanImmunogenHSP27peptideConcentration1mg/mlConjugatesAlkalinePhosphatase,APC,ATTO390,ATTO488,ATTO565,ATTO594,ATTO633,ATTO655,ATTO680,ATTO700,Biotin,FITC,HRP,PE/ATTO594,PerCP,RPE,Streptavidin,Unconjugated PerCP Overview:Peridinin-Chlorophyll-ProteinComplexSmallphycobiliproteinIsolatedfromredalgaeLargestokesshift(195nm)MolecularWeight:35kDaPerCPDatasheet OpticalProperties:λex =482nmλem=677nmεmax=1.96x106Laser =488nm  PE/ATTO594PE/ATTO594isatandemconjugate,wherePEisexcitedat535nmandtransfersenergytoATTO594viaFRET(fluorescenceresonanceenergytransfer),whichemitsat627nm.Overview:HighfluorescenceyieldHighphotostABIlityVeryhydrophilicExcellentsolubilityinwaterVerylittleaggregationPE/ATTO594DatasheetOpticalProperties:λex =535nmλem =627 nmLaser = 488to561nm  FITC(Fluorescein)Overview:ExcellentfluorescencequantumyieldHighrateofphotobleachingGoodsolubilityinwaterBroademissionspectrumpHdependentspectraMolecularformula:C20H12O5Molarmass:332.3g/molFITC-Fluorescent-conjugateOpticalProperties:λex =494 nmλem=520 nmεmax=7.3×104Φf= 0.92τfl =5.0nsBrightness= 67.2Laser = 488 nmFilterset = FITC  ATTO700Overview:HighfluorescenceyieldExcellentthermalandphotostabilityQuenchedbyelectrondonorsVeryhydrophilicGoodsolubilityinpolarsolventsZwitterionicdyeMolarMass:575g/molATTO700Datasheet OpticalProperties:λex =700 nmλem=719nmεmax=1.25×105Φf= 0.25τfl =1.6nsBrightness= 31.3Laser =676nmFilterset =Cy®5.5  ATTO680Overview:HighfluorescenceyieldExcellentthermalandphotostabilityQuenchedbyelectrondonorsVeryhydrophilicGoodsolubilityinpolarsolventsZwitterionicdyeMolarMass:631g/molATTO680Datasheet OpticalProperties:λex =680nmλem=700 nmεmax=1.25×105Φf= 0.30τfl =1.7nsBrightness=37.5Laser =633 –676nmFilterset =Cy®5.5  ATTO655Overview:HighfluorescenceyieldHighthermalandphotostabilityExcellentozoneresistanceQuenchedbyelectrondonorsVeryhydrophilicGoodsolubilityinpolarsolventsZwitterionicdyeMolarMass:634g/molATTO655DatasheetOpticalProperties:λex =663nmλem=684nmεmax=1.25×105Φf= 0.30τfl =1.8nsBrightness= 37.5Laser =633 –647nmFilterset =Cy®5  ATTO633Overview:HighfluorescenceyieldHighthermalandphotostabilityModeratelyhydrophilicGoodsolubilityinpolarsolventsStableatpH4–11Cationicdye,perchloratesaltMolarMass:652.2g/molATTO633DatasheetOpticalProperties:λex =629nmλem=657nmεmax=1.3×105Φf= 0.64τfl =3.2nsBrightness=83.2Laser =633 nmFilterset =Cy®5  ATTO594Overview:HighfluorescenceyieldHighphotostabilityVeryhydrophilicExcellentsolubilityinwaterVerylittleaggregationNewdyewithnetchargeof-1MolarMass:1137g/molATTO594Datasheet OpticalProperties:λex =601 nmλem=627nmεmax=1.2×105Φf= 0.85τfl =3.5nsBrightness=102Laser =594nmFilterset =TexasRed®  ATTO565Overview:HighfluorescenceyieldHighthermalandphotostabilityGoodsolubilityinpolarsolventsExcellentsolubilityinwaterVerylittleaggregationRhodaminedyederivativeMolarMass:611g/molATTO565Datasheet OpticalProperties:λex =563nmλem=592nmεmax=1.2×105Φf= 0.9τfl =3.4nBrightness=10Laser =532nmFilterset = TRITC  ATTO488Overview:HighfluorescenceyieldHighphotostabilityVeryhydrophilicExcellentsolubilityinwaterVerylittleaggregationNewdyewithnetchargeof-1MolarMass:804g/mol ATTO488Datasheet  OpticalProperties:λex =501nmλem=523nmεmax=9.0×104Φf= 0.80τfl =4.1nsBrightness= 72Laser = 488 nmFilterset = FITC  ATTO390Overview:HighfluorescenceyieldLargeStokes-shift(89nm)GoodphotostabilityModeratelyhydrophilicGoodsolubilityinpolarsolventsCoumarinderivate,unchargedLowmolarmass:343.42g/mol ATTO390DatasheetOpticalProperties:λex =390nmλem=479nmεmax=2.4×104Φf= 0.90τfl =5.0nsBrightness= 21.6Laser =365or405nm APC(Allophycocyanin)Overview:HighquantumyieldLargephycobiliprotein6chromophorespermoleculeIsolatedfromredalgaeMolecularWeight:105kDaAPCDatasheet OpticalProperties:λex =650nmλem=660nmεmax=7.0×105Φf= 0.68Brightness= 476Laser =594or633 nmFilterset =Cy®5 StreptavidinProperties:Homo-tetramericproteinpurifiedfromStreptomycesavidiniiwhichbindsfourbiotinmoleculeswithextremelyhighaffinityMolecularweight:53kDaFormula:C10H16N2O3SApplications:Westernblot,immunohistochemistry,andELISAStreptavidinDatasheetBiotinProperties:BindstetramericavidinproteinsincludingStreptavidinandneuravidinwithveryhighaffinityMolarmass:244.31g/molFormula:C10H16N2O3SApplications:Westernblot,immunohistochemistry,andELISABiotinDatasheetHRP(HorserADIshperoxidase)Properties:EnzymaticactivityisusedtoamplifyweaksignalsandincreasevisibilityofatargetReadilycombineswithhydrogenperoxide(H2O2)toformHRP-H2O2complexwhichcanoxidizevarioushydrogendonorsCatalyzestheconversionof:Chromogenicsubstrates(e.g.TMB,DAB,ABTS)intocolouredproductsChemiluminescentsubstrates(e.g.luminolandisoluminol)intolightemittingproductsviaenhancedchemiluminescence(ECL)Fluorogenicsubstrates(e.g.tyramine,homovanillicacid,and4-hydroxyphenylaceticacid)intofluorescentproductsHighturnoverrateenablesrapidgenerationofastrongsignal44kDaglycoproteinExtinctioncoefficient:100(403nm)Applications:Westernblot,immunohistochemistry,andELISAHRPDatasheetAP(AlkalinePhosphatase)Properties:Broadenzymaticactivityforphosphateestersofalcohols,amines,pyrophosphate,andphenolsCommonlyusedtodephosphorylatethe5’-terminiofDNAandRNAtopreventself-ligationCatalyzestheconversionof:Chromogenicsubstrates(e.g.pNPP,naphtholAS-TRphosphate,BCIP)intocolouredproductsFluorogenicsubstrates(e.g.4-methylumbelliferylphosphate)intofluorescentproductsMolecularweight:140kDaApplications:Westernblot,immunohistochemistry,andELISAAPDatasheet R-PE(R-Phycoerythrin)Overview:BroadexcitationspectrumHighquantumyieldPhotostableMemberofthephycobiliproteinfamilyIsolatedfromredalgaeExcellentsolubilityinwaterMolecularWeight:250kDaR-PEDatasheet OpticalProperties:λex =565nmλem=575nmεmax=2.0×106Φf= 0.84Brightness=1.68x103Laser = 488to561nmFilterset = TRITC PropertiesStorageBufferPBS,50%glycerol,0.09%sodiumazideStorageTemperature-20ºCShippingTemperatureBlueIceor4ºCPurificationProteinGPurifiedClonalityMonoclonalCloneNumber8A7IsotypeIgG1KappaSpecificityDetects~25kDaor~27kDa.RecognizesHSP25andHSP27,crossreactswithalphaBcrystallin.CiteThisProductStressMarqBiosciencesCat#SMC-114,RRID:AB_2120775CertificateofAnalysisA1:5000dilutionofSMC-114wassufficientfordetectionofHSP27in20µgofHeLacelllysatebyECLimmunoblotanalysis.BIOLOGicalDescriptionAlternativeNamesHSPB2Antibody,Heatshockproteinbeta-2Antibody,HSPB2Antibody,DMPK-bindingproteinAntibody,MKBPAntibodyResearchAreasCancer,HeatShockCellularLocalizationCytoplasm,Cytoskeleton,Nucleus,SpindleAccessionNumberNP_001532.1GeneID3315SwissProtP04792ScientificBackgroundHSP25isthemousehomologueofthehumanHSP27protein,amemberofthesmallheatshockproteinfamilycomprisedofadiversegroupofproteinsfrom~15to>30kDa(1).ThebasicstructureofmostsHSPsisahomologousandhighlyconservedaminoacidsequence,withanα-crystallin-domainattheC-terminusandtheWD/EPFdomainatthelessconservedN-terminus.ThisN-terminusisessentialforthedevelopmentofhighmolecularoligomers(2,3).HSP27-oligomersconsistofstabledimersformedbyasmanyas8-40HSP27proteinmonomers(4).Theoligomerizationstatusisconnectedwiththechaperoneactivity:aggregatesoflargeoligomershavehighchaperoneactivity,whereasdimershavenochaperoneactivity(5).HSP27islocalizedtothecytoplasmofunstressedcellsbutcanredistributetothenucleusinresponsetostress,whereitmayfunctiontostabilizeDNAand/orthenuclearmembrane.Itcanberapidlyphosphorylatedinresponsetophysiologicalstimulirelevanttothecelltypeexamined.Thus,HSP27hasbeensuggestedtobeanimportantintermediateinsecondmessenger-mediatedsignalingpathways(6).Otherfunctionsincludechaperoneactivity(asmentionedabove),Thermo-toleranceinvivo,inhibitionofapoptosis,andsignaltransduction.Specifically,invitro,itactsasanATP-independentchaperonebyinhibitingproteinaggregationandbystabilizingpartiallydenaturedproteins,whichensuresrefoldingoftheHSP70complex.HSP27isalsoinvolvedintheapoptoticsignalingpathwaybecauseitinterfereswiththeactivationofcytochromec/Apaf-1/dATPcomplex,therebyinhibitingtheactivationofprocaspase-9.ItisalsohypothesizedthatHSP27mayservesomeroleincross-bridgeformationbetweenactinandmyosin(7).Andfinally,HSP27isalsothoughttobeinvolvedintheprocessofcelldifferentiation.Theup-regulationofHSP27correlateswiththerateofphosphorylationandwithanincreaseoflargeoligomers.ItispossIBLethatHSP27mayplayacrucialroleinterminationofgrowth(8).FormoreinformationvisitourHSP27ScientificResourceGuideathttp://www.HSP27.com.References1.WelchW.J.(1985)JBiol.Chem.260:3058-3062.2.KimK.K.,KimR.,andKimS.(1998)Nature394(6693):595-599.3.VanMontfortR.,SlingsbyC.,andVierlingE.(2001)AddvProteinChem.59:105-56.4.EhrnspergerM.,GraberS.,GaestelM.andBuchnerJ.(1997)EMBOJ.16:221-229.5.CioccaD.R.,OesterreichS.,ChamnessG.C.,McGuireW.L.,andFuguaS.A.(1993)JNatlCancerInst.85(19):1558-70.6.WelshM.J.,WuW.,ParvinemM.,andGilmontR.R.(1996)Biol.OfReprod.55:141-151.7.SartoC.BinnzP.A.andMocarelliP.(2000)Electrophoresis.21(6):1218-26.8.ArrigoA.P.(2005)JCellBiochem.94(2):241-6.9.Jia,Y.etal.(2001)J.Biol.Chem.276(43):39911-39918.ProductImagesImmunohistochemistryanalysisusingMouseAnti-Hsp27MonoclonalAntibody,Clone8A7(SMC-114).Tissue:embryosomites.Species:Rat.PrimaryAntibody:MouseAnti-Hsp27MonoclonalAntibody(SMC-114)at1:1000.SecondaryAntibody:FITCGoatAnti-Mouse(green).Counterstain:Rhodamine-phalloidinlabeledactin(red).Courtesyof:MikeWelsh,Umich.ImmunohistochemistryanalysisusingMouseAnti-Hsp27MonoclonalAntibody,Clone8A7(SMC-114).Tissue:coloncarcinoma.Species:Human.Fixation:Formalin.PrimaryAntibody:MouseAnti-Hsp27MonoclonalAntibody(SMC-114)at1:5000for12hoursat4°C.SecondaryAntibody:BiotinGoatAnti-Mouseat1:2000for1houratRT.Counterstain:MayerHematoxylin(purple/blue)nuclearstainat200µlfor2minutesatRT.Localization:Inflammatorycells.Magnification:40x.ImmunohistochemistryanalysisusingMouseAnti-Hsp27MonoclonalAntibody,Clone8A7(SMC-114).Tissue:backskin.Species:Mouse.Fixation:Bouin’sFixativeandparaffin-embedded.PrimaryAntibody:MouseAnti-Hsp27MonoclonalAntibody(SMC-114)at1:100for1houratRT.SecondaryAntibody:FITCGoatAnti-Mouse(green)at1:50for1houratRT.Localization:Epidermis.WesternBlotanalysisofRatLungtissuelysatesshowingdetectionofHsp27proteinusingMouseAnti-Hsp27MonoclonalAntibody,Clone8A7(SMC-114).Load:15µgprotein.Block:5%blockingsolution.PrimaryAntibody:MouseAnti-Hsp27MonoclonalAntibody(SMC-114)at1:1000for2hoursatRT.SecondaryAntibody:GoatAnti-Mouse:HRPfor1houratRT.ImmunohistochemistryanalysisusingMouseAnti-Hsp27MonoclonalAntibody,Clone8A7(SMC-114).Tissue:backskin.Species:Mouse.Fixation:Bouin’sFixativeandparaffin-embedded.PrimaryAntibody:MouseAnti-Hsp27MonoclonalAntibody(SMC-114)at1:100for1houratRT.SecondaryAntibody:FITCGoatAnti-Mouse(green)at1:50for1houratRT.Localization:Epidermis.ProductCitations(1)WesternBlotTheRoleofNHE8intheRegulationofRenalProximalTubuleCalciumReabsorption.Wiebe,S.(2016)UniversityofAlberta.Dissertation.PubMedID:N/AReactivityRatApplications:WesternBlot ATTO655Overview:HighfluorescenceyieldHighthermalandphotostabilityExcellentozoneresistanceQuenchedbyelectrondonorsVeryhydrophilicGoodsolubilityinpolarsolventsZwitterionicdyeMolarMass:634g/molATTO655DatasheetOpticalProperties:λex =663nmλem=684nmεmax=1.25×105Φf= 0.30τfl =1.8nsBrightness= 37.5Laser =633 –647nmFilterset =Cy®5