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当前位置: 首页 > 产品中心 > Quantitation_Kits > AAT Bioquest/Amplite™比色钙定量试剂盒*蓝色*/36361/200试验
商品详细AAT Bioquest/Amplite™比色钙定量试剂盒*蓝色*/36361/200试验
AAT Bioquest/Amplite™比色钙定量试剂盒*蓝色*/36361/200试验
AAT Bioquest/Amplite™比色钙定量试剂盒*蓝色*/36361/200试验
商品编号: 36361
品牌: aatbio
市场价: ¥42060.00
美元价: 25236.00
产地: 美国(厂家直采)
公司:
产品分类: 定量试剂盒
公司分类: Quantitation_Kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
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Ex/Em(nm)652/None
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategorySmallMoleculeDetection
MetalIons
RelatedDiagnosticMolecules
BiochemicalAssays
Calciumisessentialforalllivingorganisms,particularlyincellphysiology,wheremovementofthecalciumionCa2+intoandoutofthecytoplasmfunctionsasasignalformanycellularprocesses.Calciumisthefifthmostabundantelementbymassinthehumanbody,whereitisacommoncellularionicmessengerwithmanyfunctions,andservesalsoasastructuralelementinbone.Calciumplaysanimportantroleinmediatingtheconstrictionandrelaxationofbloodvessels,nerveimpulsetransmission,musclecontraction,andhormonesecretion.Theserumlevelofcalciumiscloselyregulatedwithinafairlylimitedrange(9to10.5mg/dL)inthehumanbody.Bothhypocalcaemiaandhypercalcaemiaareseriousmedicaldisorders.Causesoflowcalciumlevelsincludechronickidneyfailure,vitaminDdeficiency,andlowbloodmagnesiumlevelsthatcanoccurinseverealcoholism.Amplite™CalciumDetectionKitprovidesasimplemethodfordetectingcalciuminphysiologysolutions.ThiskitusesourCalciumBlue™asthechromogeniccalciumindicator.Itsabsorbancechangesinresponsetocalciumbinding.CalciumBlue™bindscalciumtightlyintheneutralpHrange,generatingCalciumBlue™-calciumcomplexthathasintenseabsorptionat~650nm.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
AssayProtocol

1. Calciumstandardsandtestsamplepreparations:

1.1   Take10μLof300mMCalciumStandardSolution(ComponentC)to990µLDilutionBuffer(ComponentB)toget3

 mMCalciumstandardsolution.

 

1.2   Add50μLof3mMCalciumStandardsolution(fromStep1.1)to950µLDilutionBuffer(ComponentB)toget150µM  

Calciumstandardsolution.

1.3   Take500μLof150µMCalciumStandardSolution(fromStep1.2)toperform1:2serialdilutionstoget75,37.5,18.75,

9.375,4.68,2.34and0µMseriallydilutedCa2+standards.

1.4   Dilutethetestsampleto5-100µMrangewithDilutionBuffer(ComponentB).

1.5   Add50μLofCalciumStandard,dilutedsamplesandblankcontrolintoeachwellasshowninTable1.


 

Table1.Layoutofcalciumstandardsandtestsamplesinaclearbottom96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

CS1

CS1

….

….

….

….

 

 

 

 

 

 

CS2

CS2

 

 

 

 

 

 

 

 

 

 

CS3

CS3

 

 

 

 

 

 

 

 

 

 

CS4

CS4

 

 

 

 

 

 

 

 

 

 

CS5

CS5

 

 

 

 

 

 

 

 

 

 

CS6

CS6

 

 

 

 

 

 

 

 

 

 

CS7

CS7

 

 

 

 

 

 

 

 

 

 

Note:CS=CalciumStandards,BL=BlankControl,TS=TestSamples.

Table2.Reagentcompositionforeachwell

CalciumStandard

BlankControl

TestSample

Serialdilutions*(50μL)

DilutionBuffer(50μL)

50μL

*Note:Addtheseriallydilutedcalciumstandardsfrom150µMto2.34µMintowellsfromCS1toCS7induplicate.

2.  Runcalciumassay:

2.1    Add50μLofCalciumBlue™(ComponentA)toeachwellofcalciumstandards,blankcontrol,andtestsamples

(seeStep1.5,Table1and2)tomakethetotalcalciumassayvolumeto100µL/well.

Note: Fora384-wellplate,add25μLofsampleand25μLofassayreactionmixtureintoeachwell.

2.2   Incubatethereactionfor5-10minutesatroomtemperature,protectedfromlight.

2.3   Measuretheabsorbanceintensitiesat600nmor650nm.

3. Calciumcalibrationcurve:

           Plottheassaydataasshownbelow.

Graph1.jpg

 

Figure1.Calciumdoseresponsewasmeasuredona96-wellblackwall/clearbottomplatewiththeAmplite™Colorimetric

CalciumQuantitationKit.Aslowas~2.5µMCa2+wasdetectedwith5minutesincubationtime(n=3).


AssayProtocolforSerumorUrineSamples

1. Preparecalciumstandardsandtestsamplepreparations:

1.1   Take10μLof300mMCalciumStandardsolution(ComponentC)to990µLDilutionBuffer(ComponentB)toget3mMCalciumStandardSolution.

1.2   Take500μLof3mMCalciumStandardSolutiontoperform1:2serialdilutionstoget1.5,0.75,0.375,0.1875,0.094,0.047and0µMseriallydilutedCa2+standards.

1.3   Add10μLofcalciumstandard,serumorurinesamplesandblankcontrolintoeachwellasshowninTable3.

 

Table3.Layoutofcalciumstandardsandtestsamplesinaclearbottom96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

CS1

CS1

….

….

….

….

 

 

 

 

 

 

CS2

CS2

 

 

 

 

 

 

 

 

 

 

CS3

CS3

 

 

 

 

 

 

 

 

 

 

CS4

CS4

 

 

 

 

 

 

 

 

 

 

CS5

CS5

 

 

 

 

 

 

 

 

 

 

CS6

CS6

 

 

 

 

 

 

 

 

 

 

CS7

CS7

 

 

 

 

 

 

 

 

 

 

Note:CS=CalciumStandards,BL=BlankControl,TS=TestSamples.

 

Table4.Reagentcompositionforeachwell

CalciumStandard

BlankCo

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。
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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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