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当前位置: 首页 > 产品中心 > Specialty_ELISA_kits > Diapharma/TECO®超灵敏输卵管卵黄原蛋白ELISA/TE1049/Kit/96试验
商品详细Diapharma/TECO®超灵敏输卵管卵黄原蛋白ELISA/TE1049/Kit/96试验
Diapharma/TECO®超灵敏输卵管卵黄原蛋白ELISA/TE1049/Kit/96试验
Diapharma/TECO®超灵敏输卵管卵黄原蛋白ELISA/TE1049/Kit/96试验
商品编号: TE1049
品牌: Diapharma
市场价: ¥0.00
美元价: 0.00
产地: 美国(厂家直采)
公司:
产品分类: 特色ELISA试剂盒
公司分类: Specialty_ELISA_kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍

Description:

TheTECO®UltraSensitiveSalmonid VitellogeninELISAkitisaverysensitiveenzymelinkedimmunosorbentassayforthequantitativedeterminationofvitellogenin(VTG)insalmonidfishmucus orcellculture*.

Cyprinid Vitellogenin ELISA, Perch (Perciformes) Vitellogenin ELISA

VitellogenindeterminationisoneofthecoreendpointsinscreeningandtestingforendocrinedisruptingchemicalsstandardizedintheOECDGuidlinesforthetestingofchemicalsforestrogenicactivity:

  •  OECD(2009),TestNo.229
  •  OECD(2009),TestNo.230
  •  OECD(2011),TestNo.234

Vitellogenindeterminationisusedinecotoxicologicalstudiestodeterminetheeffectsofestrogeniccompoundsinthewater.

Range:0.012-1.0ng/ml

Sensitivity: <0.02ng/ml

Incubationtime: 4hours

SampleVolume: 50µl

SamplePreparation:

  • Mucus: CollectasdescribedintheTECO®MucusCollectionSet(TE1034).Mucuscontainingswabscanbestoredseveralmonthsat<-20°C.
  • Cellculture:Storefreshsamplesimmediatelyaftercollectionat<-20°Cuntilassayed.

ReferenceValues:

  • Mucuslevelsareintherangeofng/ml.

*Additionalassayvalidationmayberequiredwiththeuseofcellculturesamples.

KitComposition:

KitContents:

  • AntibodyCoatedMicroassayPlate: 96-wellplate (12×8break-apartwellstripscoatedwithIgGdirectedagainstSalmonid VTG)
  • StandardStockSolution:17.5 ng,2vials
  • ControlC1:  lowcontrol,2vials
  • ControlC2: highcontrol,2vials
  • WashBuffer: 1x30ml,50X (Dilute1:50withdeionizedwater.)
  • DilutionBuffer:1x55ml.Readytouse.
  • MatrixSolution:1x7ml.Readytouse.
  • BiotinylatedAntibody(Biotin-AB):1x12ml.Readytouse.
  • StreptavidinPeroxidaseConjugate (SA-HRPConjugate):1x12ml. Readytouse.
  • TMBSubstrate:1x12ml. Readytouse.
  • StopSolution: 1x12ml 1MHCl.Readytouse.

Materialsrequiredbut notsupplied:

Pipettes,10μl–1000μl
•Multichannelpipettes,50μl–100μl
•Graduatedcylindersforreconstitutingand dilutingreagents
•ManualAspirationSystemorautomaticwasherforELISAplates
•Distilledwater
•Vortexmixer
•ELISAplatereadersuitablefor96wellformatsandcapableofmeasuringat450nm(Reference:590-650nm)
•ELISAplateshaker(500rpm)
•Softwarepackagefordatagenerationandanalysis

Formucussamples:TheMucusCollectionSet(TE1034)whichincludesExtractionBufferandvalidatedSamplingSwabsis alsorequired.

AssayPrinciple:

AssayPrinciple

TheTECO®UltraSensitiveSalmonid VitellogeninELISAkitisa96-wellimmuno-captureELISAproduct.Mucusandcellculturesamplesareincubatedwiththevitellogeninspecificantibody-coatedmicrotiterplate.Afterunboundmaterialiswashedout,apolyclonalbiotinylated antibodybindstothevitellogenin.Inthefollowingincubationstep,astreptavidin-peroxidaseconjugatebindstothebiotinylatedantibody.Inthefinalsubstratereaction,colordevelopmentisdirectlyproportionaltotheamountofvitellogenininthesample.

ResultAnalysis

Thestandardrangeoftheassayisbetween0and1.0 ng/ml.Acalibrationcurvecanbeestablishedbyplottingstandardconcentrationonthex-axis(linearscale)againsttheabsorbanceofthestandardsonthey-axis(linearscale).Thevitellogeninconcentrationsinsamples canthenbereadoffthecalibrationcurve. A4-parametercurvefitshouldbeusedforautomaticdatareduction.Ifsampleswerepre-diluted,theconcentrationcan beobtainedbymultiplyingthevaluereadoffthecalibrationcurvebythedilutionfactor.Adilutioncorrectionformucusisnotnecessaryifonlythe0.5mlExtractionBufferwasaddedtotheswab.SampleswithhigherabsorbancevaluesthanStandardAshouldbetestedagain, pre-dilutedwithDilutionBuffer, andthenthis additionaldilutionshould betakeninaccountfortheconcentrationcalculation.

TECO®UltraSensitivSalmonid VTG ELISAStandardCurveExample

Species:

  • Atlanticsalmon(Salmosalar)
  • Browntrout(Salmotrutta)
  • Rainbowtrout(Oncorhynchusmykiss)
  • Chumsalmon(Oncorhynchusketa)
  • Pinksalmon/humpbacksalmon(Oncorhynchusgorbuscha)
  • Brooktrout(Salvelinusfontinalis)
  • Coregonusspec.

Background:

Inoviparousanimals,vitellogenin(VTG)isanestrogen-inducedyolkprecursorproteinmainlysynthesizedinthelivertobedepositedinthematuringoocytes,whereitissplitintheyolkproteinslipovitellin1,lipovitellin2andphosvitin.Theseyolkproteinsserveasnourishmentstorageforthedevelopingembryos.Non-physiologicalinductionofvitellogenininmalesorinjuvenilefishisthoughttoindicateanestrogenmediatedendocrinedisruption.Therefore,VTGdeterminationisoneofthecoreendpointsinscreeningandtestingforendocrinedisruptingchemicalsstandardizedintheOECDGuidelinesforthetestingofchemicalsforestrogenicactivity.Normally,vitellogeninismeasuredinbloodsamplesorwholebodyhomogenate(WBH)–bothsampletypesrequireinvasiveanddestructivetreatmentofthefish.Bloodcanbe difficulttocollect,inparticularwhereverysmallfishareconcernedand wheretheanimalsmustsurvivesampling.

Recently,severalcelltypeshavebeenshowntoproduceVTGafterestrogenstimulation,includingthoseoftheepidermalmucosa.EventhoughtheVTGconcentrationintheskinmucusisanorderofmagnitudelowerthaninbloodserumorinbodyhomogenates(containinglivertissue),theskinmucosaisverywellsuitedasamatrixfor determiningexogenousVTGinductioncausedbyenvironmentalchemicalswithaffinitytoestrogenreceptors.ByusingahighlysensitiveELISAincombinationwithauniquesamplingandextractionsystemthedeterminationofmucosa-bornVTGdeterminationhasthefollowingadvantages:

  • Simpleandhighlystandardizedsamplingtechniqueandsamplepreparation.
  • Strictlydefinedmatrixwithoutproteasecontaminationcausedbynon-targettissuesorlymphaticfluid.
  • Non-destructiveandtherebyallowingseveralsubsequentsamplingsinordertorecordakineticofVTGinductionwithamaximumknowntoappearafter7daysofexposure.ThereforeMucosatestarefullycompatIBLewithacuteaswellaschronicOECDtestmethods.
  • Epithelialorganizedepidermisisdirectlyexposedtoexogenousestrogenstherebyallowingadirectcomparisonwithinvitrotestusingestrogensensitivevitellogeninproducingfishcelllines.
  • LowerdegreeofinterferencewithendogenousVTGproduction(infemales)andbioconcentrationorenterohepaticcirculationoftheeffectiveestrogen(xenoestrogen)andtherebyshowingacleardoseresponserelationship.
  • StABIlityofstandardsandsamplesifprescribedstorageconditionsareobserved.

TheTECO®UltraSensitiveSalmonid VitellogeninELISAkitprovidesahigh-sensitivityELISAoptionforresearchersdeterminingVTGproteinconcentrationinfishmucusorcellculturesamples.

品牌介绍
Diapharma使命宣言位于俄亥俄州西切斯特的Difarma Group,Inc.在诊断和研究领域销售止血、血栓形成、血小板功能测试、仪器和凋亡产品,并提供强大的技术能力和经验,以确保满足或超过客户的期望。地黄止血显色凝块酶联免疫吸附试验试剂盒历史1997年1月1日,由俄亥俄州富兰克林市的Pharmacia Hepar,Inc.成立,最初是Chromogenix基质和分析的独家美国和加拿大经销商。四分之一个多世纪前,Chromogenix开发了第一个显色底物技术,其前身是Kabi Diagnostica。卡比后来与法玛西亚合并。希帕玛目前的一些员工在法玛西亚肝素制造厂的显色部门工作。1998年,夏帕玛搬到了俄亥俄州的西切斯特,至今仍在那里。多年来,迪法玛扩大了其产品线,包括各种止血、细胞死亡、血小板功能、生态毒理学、化验、试剂、抗体和高级制造商的仪器。2017年,夏帕玛庆祝了20年的成功