请使用支持JavaScript的浏览器!
主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Amplite™ Colorimetric Calcium Quantitation Kit *Blue Color*/36361/200 Tests
产品编号:36361
市  场 价:¥42060.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$2103.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Amplite™ Colorimetric Calcium Quantitation Kit *Blue Color*/36361/200 Tests
商品介绍
Overview
PrinterFriendlyVersion

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

AAT Bioquest常用产品报价单

产品名货号公司分类商城分类美金价
AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg2iFluor Rapid Tests酸碱缓冲液1088.00
AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug8B0001Signaling Intermediate Ab重组抗体2828.00
AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug8A0022Phospho-Specific Ab受体2828.00
AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug16440Fluorescent Anti-IgGs荧光染料1088.00
AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug16518Fluorescent Anti-IgGs抗小鼠4278.00
AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug16900Streptavidin Conjugates其他生物染料1378.00
AAT Bioquest/GCNT3 Antibody/8C14708/50 ug8C14708Signaling Intermediate Ab功能性抗体2828.00
AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate12518Reporter Gene Enzymes其它检测试剂盒2103.00
AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi1100mFluor™ Dyes and Kits标记试剂盒2103.00
AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests10058Diagnostic Molecules定量试剂盒2828.00
AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each38101cAMP GPCR Assays常规细胞株0.00
自营商城图标
厂家直采
全球直采 正品优价
正品保障图标
正品保障
厂家直发 有线跟踪
解放采购图标
正规清关
CIF100%正规报关,提供发票
及时交付图标
及时交付
限时必达 不达必赔