©BioservUK2017
BioServUKLtdTheInnovationCentre,217Portobello,Sheffield,S14DPTel:+44(0)1142242235Email:[email protected]
DiaserveLaboratoriesGmbHSeeshaupterStr.27,82393,Iffeldorf,GermanyTel:+49(0)8856803605Email:[email protected]
Page 1 of 4
DatasheetAnti-DAZLClone3/11AProductName AntiHumanDAZL3/11A
CatalogueNumber 3/11A
Clone,Isotype 3/11A,IgG1
Format IgG
TestedApplications WB,IHC,IF
Description:DeletedinAzoospermia-like(DAZL)proteinplaysaroleingermcelldevelopment.Mutationsinthisgenearelinkedtospermatogenicfailureandinfertilityinmales.Clone3/11ArecognizesDAZLatitsCterminus,andisusedindetectionofDAZLexpressionbyvariousmethods.ProductDetails:Forminstock:IgG,purified–1.0mg/mL.Alsoavailableasunpurifiedsupernatant.Host:MouseSpecificity:RecognizesDAZLproteinCterminussequenceCRVHHFRRSRAMLKSV.HumanHistologypositivecontrol:OvariesorTestisFusionpartner:SpleencellsfromimmunisedT/OoutbredmicewerefusedwithcellsoftheSP2/0myelomacellline.Storage:Storeat+4°Cor-20°C.Avoidrepeatedfreezingandthawing.Shelflife:18monthsfromdateofdispatch.Regulatory/Restrictions:Forresearchandcommercialpurposes.Applications SuggestedDilutionWesternBlot 1:100-1:1000Immunohistochemistry-Paraffin 1:50-1:500Immunofluorescence 1:50-1:500
©BioservUK2017
BioServUKLtdTheInnovationCentre,217Portobello,Sheffield,S14DPTel:+44(0)1142242235Email:[email protected]
DiaserveLaboratoriesGmbHSeeshaupterStr.27,82393,Iffeldorf,GermanyTel:+49(0)8856803605Email:[email protected]
Page 2 of 4
Applications:
Clone3/11AusedtodetectDAZLexpressioninhumanovarycellsbyImmunofluorescenceImagecaption:...(B)TriplefluorescentimmunohistochemistryforGDF9(green),DAZL(blue)andBOLL(red)in20weekhumanfetalovarywithDAPIascounterstain(grey).Splitchannelandmergedimagesin(A)and(B)areshownasaremergedimagesofnon-immuneserumnegativecontrol(NEG).Scalebarsare20μm.(C)NucleardiametersofDAZL,BOLLandGDF9stainedgermcellsindicatesthatGDF9positivecellsaresignificantlylarger(p<0.001)thanDAZLbutnotBOLLexpressingcells(barsindicatemean±sem).(D)HighermagnificationmergedimageofGDF9/DAZL/BOLLimmunohistochemistryshowingonelargeprimordialfollicleispositiveforbothGDF9andDAZLbutotherfolliclesarepositiveonlyforDAZL.(Bayne,Retal.)Dilutionused:1:400
Clone3/11AusedtodetectDAZLinovarianandtesticulartissuebyWesternBlotImagecaption:Westernanalysisof1stand2ndtrimesterovariesandtestes.Inbothovarian(A)andtesticular(B)samplesVASA(76Kd)wasnotdetectableinthe1sttrimestersamplesbutwaspresentinthosefromthe2ndtrimester.DAZL(33Kd)waslow/undetectablein1sttrimesterovaries(C)whereasitwasdetectableinovariansamplesfrom2ndtrimesterandtesticularsamplesfromboth1stand2ndtrimester(D).OCT4(42Kd)waspresentinbothovaries(E)andtestes(F)duringboththe1stand2ndtrimester.Theloadingcontrolinallcaseswasβ-tubulin(51Kd).(Anderson,Retal.)Dilutionused:1:500
©BioservUK2017
BioServUKLtdTheInnovationCentre,217Portobello,Sheffield,S14DPTel:+44(0)1142242235Email:[email protected]
DiaserveLaboratoriesGmbHSeeshaupterStr.27,82393,Iffeldorf,GermanyTel:+49(0)8856803605Email:[email protected]
Page 3 of 4
Clone3/11AusedtodetectDAZLexpressioninhumanovarycellsbyImmunofluorescenceImagecaption:...DAZLpositivegermcellswererareinthe1sttrimester(e,ovary61d;g,testis64d)butgroupsofcells('nests',labelledN)withcytoplasmicstainingwerepresentinthe2ndtrimesterovaries...(Anderson,Retal.)Dilutionused:1:500
Clone3/11AusedtodetectDAZLexpressioninrattestisbyIHC-PImagecaption:Immunohistochemistryshowinglocalisationofgermcellsand5mCduringmidtolategestation.Inordertoclearlyshowthelocalisationofgermcellsatallstagesofdevelopment...usingDAZLasacytoplasmicmarker....(Rose,Cetal.)Dilutionused:1:500
©BioservUK2017
BioServUKLtdTheInnovationCentre,217Portobello,Sheffield,S14DPTel:+44(0)1142242235Email:[email protected]
DiaserveLaboratoriesGmbHSeeshaupterStr.27,82393,Iffeldorf,GermanyTel:+49(0)8856803605Email:[email protected]
Page 4 of 4
Clone3/11AusedtodetectDAZLexpressionincynomolgusmonkeytestesbyImmunofluorescenceonparaffintissuesectionsImagecaption:...(D–Q)ExpressionofVASA,DAZL,andSCP1proteinsduringspermatogenesisincynomolgusmonkeys...(Yamauchi,Ketal.)Dilutionused:1:200
Clone3/11AusedtodetectDAZLexpressioninovariontissuebyImmunofluorescenceImagecaption:AbnormalgermcellproliferationwasnotedinGR-deficientembryosaftersexdetermination.(Chen,SRetal.)Dilutionused:1:400
References:
1. Bayne,R.A.L.,Kinnell,H.L.,Coutts,S.M.,He,J.,Childs,A.J.,Anderson,R.A.(2015)GDF9isTransientlyExpressedinOocytesbeforeFollicleFormationintheHumanFetalOvaryandisRegulatedbyaNovelNOBOXTranscript.PLoSONE,10(3),e0119819.
2. Anderson,R.A.,Fulton,N.,Cowan,G.,Coutts,S.,Saunders,P.T.(2007).ConservedanddivergentpatternsofexpressionofDAZL,VASAandOCT4inthegermcellsofthehumanfetalovaryandtestis.BMCDevelopmentalBiology,7,136.
3. Rose,C.M.,VandenDriesche,S.,Sharpe,R.M.,Meehan,R.R.,Drake,A.J.(2014)DynamicchangesinDNAmodificationstatesduringlategestationmalegermlinedevelopmentintherat.Epigenetics&Chromatin,7,19.
4. Yamauchi,K.,Hasegawa,K.,Chuma,S.,Nakatsuji,N.,Suemori,H.(2009)InVitroGermCellDifferentiationfromCynomolgusMonkeyEmbryonicStemCells.PLoSONE,4(4),e5338.
5. Chen,S-R.,Zheng,Q-S.,Zhang,Y.,Gao,F.,Liu,Y-X.(2013)Disruptionofgenitalridgedevelopmentcausesaberrantprimordialgermcellproliferationbutdoesnotaffecttheirdirectionalmigration.BMCBiology,11,22.
