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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Cell Meter™ Fluorimetric Phagocytosis Assay Kit *Red Fluorescence*/21225/100 Tests
商品详细AAT Bioquest/Cell Meter™ Fluorimetric Phagocytosis Assay Kit *Red Fluorescence*/21225/100 Tests
AAT Bioquest/Cell Meter™ Fluorimetric Phagocytosis Assay Kit *Red Fluorescence*/21225/100 Tests
AAT Bioquest/Cell Meter™ Fluorimetric Phagocytosis Assay Kit *Red Fluorescence*/21225/100 Tests
商品编号: 21225
品牌: aatbio
市场价: ¥85560.00
美元价: 51336.00
产地: 美国(厂家直采)
公司:
产品分类: 其它检测试剂盒
公司分类: Other_kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
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Ex/Em(nm)575/597
MWN/A
CAS#N/A
SolventN/A
StorageR/L
CategoryCellBIOLOGy
pHandIonIndicators
RelatedCellMetabolism
Phagocytosisisdefinedasthecellularuptakeofparticleswithinaplasma-membraneenvelopebyacell.TheprocessofphagocytosisiscriticalintheinnateimmuneresponsebyengulfmentanddestructionofinvADIngmicroorganisms.Phagocytosisisalsorequiredinmaintainingtissuehomeostasisandremodelingbytheclearanceofapoptoticbodies.Theuptakemechanismofphagocytosisdependsonthesizeoftheparticles,receptor-ligandinteractions,andinvolvementoftheCytoskeleton.Onceinternalized,thephagosomefuseswithlysosomestoformsecondaryphagolysosomefordigestion,resultinginprogressivedecreaseofpH.OurCellMeter™FluorimetricPhagocytosisAssayKitutilizesauniquepHdependentProtonex™600Red-latexbeadconjugates.ThebeadsareinreadytousesUSPension.Unlikemostoftheexistingfluorescentdyes,theProtonex™600Red-latexbeadconjugateisnon-fluorescentoutsideofthecells.However,itsfluorescencedramaticallyincreasesastheyareinsidetheacidicphagosomes/phagolysosomes.Thischaracteristicmakesiteasytousewithouttrypanbluequenchingstepandarobusttooltostudyphagocytosisanditsregulations.CellMeter™FluorimetricPhagocytosisAssayKitalsoincludesagreenfluorescentcellviABIlitydye,whichallowingthesimultaneousdetectionofbothlivecellsandtheprocessofphagocytosisbyfluorescentmicroscopy.Thisassaycanalsobeadaptedforfluorescencemicro-platereaderandflowcytometrydetections.
SpectrumAdvancedSpectrumViewer



Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. Preparecells:
    1. Foradherentcells:Platecellsovernightingrowthmediumat20,000-50,000cells/well/100µLfora96-wellplate.
      Note1:Eachcelllineshouldbeevaluatedontheindividualbasistodeterminetheoptimalcelldensity.
      Note2:ForRAW264.7cellsthatweusedinthisassaywerecommendtoplatecellsat50,000cells/well/100µLin96-wellplateforovernight.
  2. Preparestocksolutions:
    1. Prepare6XCytochalasinD:Add18µLof10mMCytochalasinD(finalconcentrationinthewellis10µM)or18µLofDMSOin3mLcellgrowthmedium.
    2. Prepare12XProtonex™600Red-LatexBeadsConjugatesolution:Add8µLofProtonex™600Red-LatexBeadsConjugate(ComponentA)in2mLcellgrowthmedium(containing10%FBS).
      Note:Theunusedbeadscanbestoredat4°C.
    3. Prepare12XCytoTrace™Green:Add20µLofDMSO(ComponentC)intothevialofCytoTrace™Green(ComponentB)tomake400Xstocksolution.Andthenadd5µLin2mLcellgrowthmediumtohave12XCytoTrace™Greensolution
      Note:TheunusedCytoTrace™GreenDMSOstocksolutioncanbealiquotedintosingleusevialsandstoredat-20°C.
  3. Phagocytosisassayprotocol:
    1. Add25µLof6XCytochalasinD(fromstep2.1)orDMSOsolution(fromstep2.1)intoeachwell.
      Note:TheconcentrationofCytochalasinDusedintheassayshouldbeoptimizedforeachindividualcellline.
    2. Incubatetheplateinthecellincubatorfor30minutes.
    3. Add12.5µLof12XProtonex™600Red-LatexBeadsConjugatesolution(fromstep2.2)intoeachwell.
    4. Incubatetheplateinthecellincubatorfor4hours.
      Note:Theincubationtimeshouldbeoptimizedbyusersforeachindividualcelllines.
    5. Add12.5µLof12XCellTracker(fromstep2.3)intoeachwell.
    6. Incubatetheplateinthecellincubatorfor30minutes.
    7. Washtheplatetwicewith1XPBS.
  4. ObservephagocytosisinsidethecellswithTexasRedfilter(Ex/Em=570/600nm)andCytoTrace™GreenwithFITCfilter(Ex/Em=490/525nm)
References&Citations
CitationExplorer

PHD2isaregulatorforglycolyticreprogramminginmacrophages
Authors:AnnemarieGuentsch,AngelikaBeneke,LijaSwain,KatjaFarhat,ShunmugamNagarajan,BenWielockx,KaaminiRaithatha,JanDudek,PeterRehling,AnkeZieseniss
Journal:MolecularandCellularBiology(2016):MCB--00236


品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。 1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物; 2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞; 3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类; 4)致力于开发用于信号转导研究的试剂; 5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
品牌分类
淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
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