Overview:ProductNameHSF1AntibodyDescriptionRatAnti-MouseHSF1MonoclonalIgG1SpeciesReactivityHuman,Monkey,Mouse,Rat,Bovine,GuineaPig(Caviaporcellus),Hamster,RabbitApplicationsWB,IHC,ICC/IF,IP,ELISA,GSAntibodyDilutionWB(1:1000),IHC(1:8000),ICC/IF(1:100);optimaldilutionsforassaysshouldbedeterminedbytheuser.HostSpeciesRatImmunogenSpeciesMouseImmunogenPurifiedrecombinantmouseHSF1proteinConcentration1mg/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 PropertiesStorageBufferPBSpH7.4,50%glycerol,0.1%sodiumazideStorageTemperature-20ºCShippingTemperatureBlueIceor4ºCPurificationProteinGPurifiedClonalityMonoclonalCloneNumber10H4IsotypeIgG1SpecificityDetects~85kDa(unstressedcelllysates)and~95kDa(heatshockedcelllysates).CiteThisProductStressMarqBiosciencesCat#SMC-476,RRID:AB_2702279CertificateofAnalysis1µg/mlofSMC-476wassufficientfordetectionofHSF1in20µgofheatshockedHeLacelllysatebycolorimetricimmunoblotanalysisusingRabbitanti-ratIgG:APasthesecondaryantibody.BIOLOGicalDescriptionAlternativeNamesHSTF1Antibody,Heatshockfactorprotein1Antibody,Heatshocktranscriptionfactor1Antibody,HSF1AntibodyResearchAreasCancer,HeatShock,CardiovascularSystem,CellSignaling,Epigenetics,EpigeneticsandNuclearSignaling,HeartCellularLocalizationCytoplasm,NucleusAccessionNumberNP_032322.1GeneID15499SwissProtP38532ScientificBackgroundHSF1,orheatshockfactor1,belongstoafamilyofHeatShocktranscriptionfactorsthatactivatethetranscriptionofgenesencodingproductsrequiredforproteinfolding,processing,targeting,degradation,andfunction(2).Theup-regulationofHSP(heatshockproteins)expressionbystressorsisachievedattheleveloftranscriptionthroughaheatshockelement(HSE)andatranscriptionfactor(HSF)(3,4,5).MostHSFshavehighlyconservedaminoacidsequences.OnallHSFsthereisaDNAbindingdomainattheN-terminus.Hydrophobicrepeatslocatedadjacenttothisbindingdomainareessentialfortheformationofactivetrimers.TowardstheC-terminalregionanothershorthydrophobicrepeatexists,andisthoughttobenecessaryforsuppressionoftrimerization(6).Therearetwomainheatshockfactors,1and2.MouseHSF1existsastwoisoforms,howeverinhighereukaryotesHSF1isfoundinadiffusecytoplasmicandnucleardistributioninun-stressedcells.Onceexposedtoamultitudeofstressors,itlocalizestodiscretenucleargranuleswithinseconds.Asitrecoversfromstress,HSF1dissipatesfromthesegranulestoadiffusenuceloplasmicdistribution.HSF2ontheotherhandissimilartomouseHSF1,asitexistsastwoisoforms,thealphaformbeingmoretranscriptionallyactivethanthesmallerbetaform(7,8).VariousexperimentshavesuggestedthatHFS2mayhaverolesindifferentiationanddevelopment(9,10,11).References1.CottoJ.J.,FoxS.G.andMorimotoR.I.(1997)J.CellScience110:2925-2934.2.MoranoK.A.andThieleD.J.(1999).GeneExpression7(6):271-82.3.TanakaK.I.,etal.(2007).JBCPapersOnlineManuscriptM704081200.4.MorimotoR.I.(1998)GenesDev12:3788-3796.5.McMillanD.R.,etal.(1998)JBioChem273:7523-7528.6.JollyC.,UssonY.andMorimotoR.I.(1999)Proc.Natl.Acad.Sci.USA96(12):6769-6774.7.FiorenzaM.T.,etal.(1995)NucleicAcidsRes.23(3):467-474.8.GoodsonM.L.,Park-SargeO.K.andSargeK.D.(1995)Mol.Cell.Biol.15(10):5288-5293.9.RalluM.,etal.(1997)Proc.Natl.Acad.Sci.USA94(6):2392-2397.10.SargeK.D.,etal.(1994)Biol.Reprod.50(6):1334-1343.11.MurphyS.P.,GorzowskiJ.J.,SargeK.D.andPhillipsB.(1994)Mol.Cell.Biol.14(8):5309-5317.ProductImagesImmunocytochemistry/ImmunofluorescenceanalysisusingRatAnti-HSF1MonoclonalAntibody,Clone10H4(SMC-476).Tissue:HeatShockedHeLaCells.Species:Human.Fixation:2%Formaldehydefor20minatRT.PrimaryAntibody:RatAnti-HSF1MonoclonalAntibody(SMC-476)at1:100for12hoursat4°C.SecondaryAntibody:R-PEGoatAnti-Rat(yellow)at1:200for2hoursatRT.Counterstain:DAPI(blue)nuclearstainat1:40000for2hoursatRT.Localization:Cytoplasm.Localizestothenucleusuponactivation.Magnification:100x.(A)DAPI(blue)nuclearstain.(B)Anti-HSF1Antibody.(C)Composite.HeatShockedat42°Cfor1h.ImmunohistochemistryanalysisusingRatAnti-HSF1MonoclonalAntibody,Clone10H4(SMC-476).Tissue:Breastcarcinoma.Species:Human.Fixation:10%FormalinSolutionfor20hoursatRT.PrimaryAntibody:RatAnti-HSF1MonoclonalAntibody(SMC-476)at1:8000for40min.SecondaryAntibody:DakolabeledPolymerHRPAnti-ratIgG,DABChromogen(brown)(DakoEnvision+System)for30minatRT.Counterstain:Mayer’sHematoxylin(purple/blue)nuclearstainfor1minuteatRT.Localization:Nuclear.Magnification:100X.Courtesyof:Dr.SandroSantagata,HarvardMedicalSchool.WesternBlotanalysisofHumanA431andHEK293celllysatesshowingdetectionofHSF1proteinusingRatAnti-HSF1MonoclonalAntibody,Clone10H4(SMC-476).PrimaryAntibody:RatAnti-HSF1MonoclonalAntibody(SMC-476)at1:1000.Immunocytochemistry/ImmunofluorescenceanalysisusingRatAnti-HSF1MonoclonalAntibody,Clone10H4(SMC-476).Tissue:HeatShockedHeLaCells.Species:Human.Fixation:2%Formaldehydefor20minatRT.PrimaryAntibody:RatAnti-HSF1MonoclonalAntibody(SMC-476)at1:100for12hoursat4°C.SecondaryAntibody:FITCGoatAnti-Rat(green)at1:200for2hoursatRT.Counterstain:DAPI(blue)nuclearstainat1:40000for2hoursatRT.Localization:Cytoplasm.Localizestothenucleusuponactivation.Magnification:20x.(A)DAPI(blue)nuclearstain.(B)Anti-HSF1Antibody.(C)Composite.HeatShockedat42°Cfor1h.ProductCitations(0)Currentlytherearenocitationsforthisproduct. ATTO680Overview:HighfluorescenceyieldExcellentthermalandphotostabilityQuenchedbyelectrondonorsVeryhydrophilicGoodsolubilityinpolarsolventsZwitterionicdyeMolarMass:631g/molATTO680Datasheet OpticalProperties:λex =680nmλem=700 nmεmax=1.25×105Φf= 0.30τfl =1.7nsBrightness=37.5Laser =633 –676nmFilterset =Cy®5.5