Description:
TECO®ADIponectinHighSensitiveELISA.Adiponectinisa30kDaproteinanditspercentageinserumproteinsis0.01%.Invivo,itappearswithdifferentoligomersanditismainlysynthesizedbyadipocytes.Untilnow,IGF-1istheonlyknownnaturalinductorofsynthesis.
Adiponectincanbeuseintheresearchof insulinresistanceandmetabolicsyndromeaswellresearchin increasedriskoftype2diabetesmellitusandcardiovasculardisease.
Today,Adiponectinisthoughttoactasanendogenicinsulinsensitizerbydecreasingexcessiveglucoselevelswithoutincreasinginsulinconcentrationsandbystimulatingtheburningofadiposetissueinmuscleandliver.
Adiponectinisassociatedwithglucoseandlipidmetabolismandisassumedtohavedirectantiatherogeniccharacteristics.
Research significance:
•Obesity
•Atherosclerosis
•Energymetabolism
•Coronaryarterydisease
•Metabolicsyndrome
•Polycysticovarysyndrome(PCOS)
KitComposition:
ReagentsSupplied
Symbol | Description | Format |
1 | AntibodyCoatedWells 12breakapartstripsof8wells(12x8intotal), inaframe,Readytouse | 1plate |
A | StandardA 2ng/ml–lyophilized | 1x0,75ml |
B | StandardB 10ng/ml–lyophilized | 1x0,75ml |
C | StandardC 30ng/ml–lyophilized | 1x0,75ml |
D | StandardD 70ng/ml–lyophilized | 1x0,75ml |
E | StandardE 100ng/ml–lyophilized | 1x0,75ml |
L | Control1 ng/ml–lyophilized,Rangeasindicatedondatasheet | 1x0,5ml |
H | Control2 ng/ml–lyophilized,Rangeasindicatedondatasheet | 1x0,5ml |
2 | DilutionBuffer Readyforuse | 1x125ml |
3 | AntibodyHRP-Conjugate Readyforuse | 1x12ml |
4 | TMBSubstrate Readyforuse | 1x12ml |
4 | WashBuffer 20timesconcentrated | 1x50ml |
6 | StopSolution–0,2MH2S04 0,2Msulfuricacid,readyforuse | 1x12 ml |
7 | CoverforMicrotiterplate,adhesive | 2pieces |
I | Kitinstruction | 1x |
Materialsrequiredbutnotsupplied
- Pipettescapableofdispensing5µl,40µl,80µL,100µl,320µl,350µl,500µl,560µland750µl
- Graduatedcylindersforreconstitutingordilutingreagents
- ManualAspirationSystemandmulti-channelpipetteorautomaticwasher
- Aquadest
- Vortexmixer
- ELISAplatereadersuitablefor96wellformatsandcapableofmeasuringat450and405nm(Reference:590–650nm)
- ELISAplateshaker(≥400rpm)(orbitalshaker)
- Softwarepackagefordatagenerationandanalysis
AssayPrinciple&Procedure:
AssayPrinciple
TheTECO®assaykitforAdiponectinisaso-calledSandwich-Assayusingtwospecificandhighaffinitymonoclonalantibodies.TheAdiponectininthesamplesbindstothefirstantibodycoatedonthemicrotiterplate.InthefollowingstepthesecondspecificantiAdiponectin-antibodybindsinturntotheimmobilizedAdiponectin.Thesecondantibodyisbiotinylatedandwillbeincubatedinamixturewithastreptavidin-peroxidase-enzymeconjugate.IntheclosingsubstratereactiontheturnofthecolorwillbecatalyzedquantitativelydependingontheAdiponectin-levelofthesamples.
AssayPrinciple
Alldeterminations(standards,dilutedserumcontrolsanddilutedsamples)shouldbeassayed induplicate.Whenperformingtheassay,thestandards,controlseraandsamplesshouldbepipetted asfastaspossIBLe(<15minutes).
Toavoiddistortionsduetodifferencesinincubationtimes,HRPconjugate,substratesolutionand stopsolutionshouldbeaddedtotheplateinthesameorderandwiththesametimeintervalas thesamples.
Allowallreagentstostandatroomtemperature(20–25°C)foratleast30minutes.Afterreconstitution, keepthereagentsatroomtemperaturefor15minutesandthenmixgentlybeforeuse.
- Preparetheframeandtherequirednumberofcoatedstrips 1.
- Allocatethewellsofthemicrotiterplateforstandards,controlsandsamples.Dilutesamples(seekitinstructionspage11)of andcontrols(1:300).
- Pipette100µldilutionbuffer 2 induplicateintowells(blank).
- Pipette100µlofeachstandards( A till E ),dilutedControlsera( L and H )andsamples (seedilutionprotocol)intothecorrespondingwells.
- Coverthewellswithsealingtapeandincubatetheplatefor1houratroomtemperature (20–25°C)onaplateshaker(≥400–500rpm).
- Afterincubation,aspiratethewellsbyusingaplatewasherormanuallydecantbyinverting theplate.Washthewells3xwith350mldilutedwashingbuffer(15secondsincubationper cycle).Afterthelastwashcycletaptheinvertedwellsgentlyonadryabsorbentsurfaceto removeexcesswashsolution.
- Pipette100µloftheantibodyHRPconjugate 3 ineachwell.
- Coverthewellswithsealingtapeandincubatetheplatefor30minutesatroomtemperature (20–25°C)onaplateshaker(≥400–500rpm).
- Washthewells3timeswithwashingbufferasdescribedinstep6.
- Pipette100µloftheTMBsubstratesolution 4 ineachwell.
- Incubatetheplatefor15minutes,inthedark,atroomtemperature(20–25°C).
- Add100µlofstopsolution E ineachwell.
- Measurethecolorreactionwithin30minutesat450nm(referencefilterbetween 590–650nm).Withstrongcolorreactione.g.>3ODalsomeasureat405nm.
ProtocolsforthedifferentautomaticELISAsystemsareavailable.
Range&Sensitivity:
Range
1–100ng/mlnativeAdiponectin
Sensitivity
<0.6ng/ml
Sample:
Samplevolume
5µl(dilute>1:300serumandplasma).ForotherBIOLOGicalfluidsseeprotocol.
Sampletype
Serum,heparinplasma,breastmilk,urine,saliva,CSF,cellculture
Samplepreparation
- Bloodcollection–fastingisrecommended.
- Samplesarestableformaximum2daysatroomtemperature.Long-termstoragestableformaximum2yearsat-20°C.
- Max.5freezeandthawcycles.
Incubationtime
2hours
Species
Human
Referencevalues:
mg/l | |
MenAdult | 8-10 |
FemaleAdult | 10-12 |
cut-off | 10 |
Comprehensivereferencedatarelatedtoageandgenderareavailableforthistest.
Background:
Adiponectinisa30kDaprotein,presenting0,01%ofserumproteins.Itismainlysynthesized byadipocytes,butmusclecellsandhepatocyteshavealsotheABIlitytosynthesizeAdiponectin. Untilnow,IGF-Iistheonlyknownnaturalinductorofthesynthesis.ItconsistsofaCollagenlike N-terminalandaglobularC-terminaldomain.InvivoAdiponectinappearswithdifferent oligomers.Besidethetrimerandditrimer,highmolecularmultimersalsoexist.Uptonow twodifferentreceptorsareknown,bothreceptorsareubiquitaryexpressed,thoughthe distributioninthetissuesvaries.TheAdiponectinReceptor1(AdipoR1)issynthesized especiallyinmuscle-andAdipoR2inlivertissue.
Aspecialsignificanceforhighmolecularmultimers,asdescribedrepeatedly,could notbeproveninacomparativestudyofthreetestsystems.
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2.NaHCO3,Ba(OH)3,H2SO4
3.HCL,NaAlO2,NaHSO4
4.Ca(OH)2,Na2CO3,BaCO3
谢谢了
要原因
请问有什么方法检测细胞内活性氧更好,各位有什么经验,及相关试剂盒推荐一下。
A.H2SO4Na2SO4AgNO3BaCl2.
B.NaOHNa2CO3NaHSO4MgCl2
C.CaCl2NaNO3MgSO4BaCl2
D.HNO3KOHKClK2CO3
请问下有无同学需要H37RA的?我是做EAE模型的,上个月购买了BDDifco公司的H37RA(货号),因为购买的时候只能整盒6支购买,但我们用不了那么多,所以想问问有无同学需要的,100mg/支,800元/支或用等价试剂交换。地址广州。有需要的请私信,谢谢!
(1)优级纯试剂 亦称保证试剂,为一级品,纯度高,杂质极少,主要用于精密分析和科学研究,常以GR表示。
(2)分析纯试剂 亦称分析试剂,为二级品,纯度略低于优级纯,杂质含量略高于优级纯,适用于重要分析和一般性研究工作,常以AR表示。
(3)化学纯试剂 为三级品,纯度较分析纯差,但高于实验试剂,适用于工厂、学校一般性的分析工作,常以CP表示。
(4)实验试剂 为四级品,纯度比化学纯差,但比工业品纯度高,主要用于一般化学实验,不能用于分析工作,常以 LR表示。
以上按试剂纯度的分类法已在我国通用。根据化学工业部颁布的“化学试剂包装及标志”的规定,化学试剂的不同等级分别用各种不同的颜色来标志,见表1。
表1 我国化学试剂的等级及标志
bhclna2so4nano3na2co3
chclnaohna2co3nacl
dba(oh)2nahco3alcl3nahso4