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公司介绍
Mission StatementDiapharma Group, Inc. in West Chester, Ohio sells hemostasis, thrombosis, platelet function testing, instrumentation, and apoptosis products in the diagnostic and research fields and provides strong technical competence and experienc...
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Diapharma/TECO® Leptin ELISA/TE1016/Kit/96 tests

Diapharma/TECO® Leptin ELISA/TE1016/Kit/96 tests
  • Diapharma/TECO® Leptin ELISA/TE1016/Kit/96 tests
商品介绍

Description:

TECO®Leptin.Leptin,theproductoftheobgene,isarecentlydiscoveredproteohormone.ItisalmostexclusivelyproducedbydifferentiatedADIpocytesandisthoughttoplayakeyroleintheregulationofbodyweight.Leptinhasaninfluenceonthecentralnervoussystem,mainlyonthehypothalamus,bysuppressingfoodingestionandincreasingenergyconsumption.

Leptincanbeusedintheresearchof reproductionandanumberofmetabolicandendocrineaxes.


Duetoitspleiotropiceffects,leptinisavaluableresearchtoolwithregardto:

•Metabolicsyndromestudies
•Adiposityresearch
•Cachexiaandothermetabolicstudies
•Eatingdisorderresearch

KitComposition:

ReagentsSupplied

SymbolDescriptionFormat
 1 AntibodyCoatedWells
12breakapartstripsof8wells(12x8intotal), inaframe,Readytouse
1plate
 A StandardA
1ng/ml–lyophilized
1x0.75ml
 B StandardB
10ng/ml–lyophilized
1x0.75ml
 C StandardC 
25ng/ml–lyophilized
1x0.75ml
 D StandardD 
50ng/ml–lyophilized
1x0.75ml
 E StandardE
100ng/ml–lyophilized
1x0.75ml
 L Control1
lyophilized,Rangeasindicatedondatasheet
1x0.5ml
 H Control2
lyophilized,Rangeasindicatedondatasheet
1x0.5ml
 2 DilutionBuffer
Readyforuse
1x25ml
 3 Antibody-HRP-Conjugate
Readyforuse
1x12ml
 4 TMBSubstrate
Readyforuse
1x12ml
 5 WashBuffer
20timesconcentrated
1x50ml
 6 StopSolution–0.2MH2S04
0.2Msulfuricacid,readyforuse
1x12ml
 7 CoverforMicrotiterplate,adhesive2pieces
 I Kitinstruction1x

MaterialsRequiredandnotSupplied

  • Pipettescapableofdispensing20µl,100µl,350µl,500µland750µl
  • Graduatedcylindersforreconstitutingordilutingreagents
  • Manualaspirationsystemandmulti-channelpipetteorautomaticwasherforELISAplates
  • Aquadest
  • Vortexmixer
  • ELISAplatereadersuitablefor96wellformatsandcapableofmeasuringat450nm(Reference:590–650nm).
  • ELISAplateshaker(≥350rpm)(orbitalshaker)
  • Softwarepackagefordatagenerationandanalysis

AssayPrinciple&Procedure:

AssayPrinciple

TheTECO®assaykitforhumanleptinisaso-calledsandwich-assayusingtwospecificandhighaffinitymonoclonalantibodies.Theleptininthesamplesbindstothefirstantibodycoatedonthemicrotiterplate.Inthefollowingstepthesecondanti-Leptin-Antibodybindsinturntotheimmobilizedleptin.Thesecondantibodyisbiotinylatedandwillbeincubatedinamixturewithastreptavidin-peroxidase-enzymeconjugate.Intheclosingsubstratereactiontheturnofthecolorwillbecatalyzedquantitativelydependingontheleptin-levelofthesamples.


AssayProcedure

Alldeterminations(standards,controlseraandsamples)shouldbeassayedinduplicate.When performingtheassay,thestandards,controlseraandthesamplesshouldbepipettedasfast aspossIBLe(<15minutes).

Toavoiddistortionsduetodifferencesinincubationtimes,HRPconjugate,substratesolution andstopsolutionshouldbeaddedtotheplateinthesameorderandwiththesametime intervalasthesamples.

Allowaallreagentstostandatroomtemperature(20–25°C)foratleast30minutes.

  1. Preparetheframeandtherequirednumberofcoatedstrips .
  2. Allocatethewellsofthemicrotiterplateforstandards,controlsandsamples.
  3. Pipette100µldilutionbuffer 2  intoallwells.
  4. Add20µldilutionbufferinduplicate(Blank).
  5. Pipette20µlofeachstandard( A till E ),controlsera( L and H )andsamplesintothe correspondingwells.
  6. Coverthewellswithsealingtapeandincubatetheplatefor1houratroomtemperature onaplateshaker(≥350rpm).
  7. Afterincubation,aspiratethewellsbyusingaplatewasherormanuallydecantbyinverting theplate.Washthewells3xwith350µldilutedwashingbuffer(15secondsincubationper cycle).Afterthelastwashcycletaptheinvertedwellsgentlyonadryabsorbentsurfaceto removeexcesswashsolution.
  8. Pipette100µloftheantibodyHRPconjugateineachwell.
  9. Coverthewellswithsealingtapeandincubatetheplatefor30minutesatroomtemperature onaplateshaker(≥350rpm).
  10. Afterincubationwashthewells3timeswithwashingbufferasdescribedinstep7.
  11. Pipette100µloftheTMBsubstratesolution 4  ineachwell.
  12. Incubatetheplatefor15minutes,inthedark,atroomtemperature(20–25°C).
  13. Add100µlofstopsolution  6 ineachwell.
  14. Measurethecolorreactionwithin30minutesat450nm(referencefilterbetween 590–650nm).Withstrongcolorreactione.g.>3OD,alsomeasureat405nm.

ProtocolsforthedifferentautomaticELISAsystemsareavailable.

Range&Sensitivity:

Range

1–100ng/ml,recombinantleptinWHONIBSC97/594


Sensitivity

LLOD<0.25ng/ml
LLOQ1ng/ml

Sample:

Incubationtime

2hours


Samplevolume

20µl


Sampletype

Serum,heparinandEDTAplasma,urine,saliva,cellculture.


Samplepreparation

  • Normalfoodintakerhythmprovided,samplesshouldbecollectedtill2p.m.Leptinshowsamoderatecircadianvariationwithapeakat2a.m.,theleptinvaluesatthattimeareabout30to100%higher.
  • Thisvariationtogetherwiththeinfluenceoffoodintakeneedstobetakenintoaccountwhenbloodsamplesarecollected.Wholebloodshouldberefrigeratedassoonaspossiblefollowingcollection.
  • Samplesarestableformaximal2daysatroomtemperature.
  • Long-termstoragestableformaximal2yearsat-20°C.
  • Max.5freezeandthawcycles.

Species

Human

Referencevalues:

Leptinlevelsdependonageandgenderandmustbereferredtothepercentagebodyfat(suchasBMI).

Comprehensiveclinicalreferencedataareavailableforthistest.

Background:

Leptin,theproductoftheobgeneisarecentlydiscovered(1994)single-chainproteohormone withamolecularweightof16kD,whichisthoughttoplayakeyroleintheregulation ofbodyweight.Itsaminoacidsequenceexhibitsnomajorhomologieswithotherproteins. Leptinisalmostexclusivelyproducedbydifferentiatedadipocytes.Itactsonthecentral nervoussystem,inparticularthehypothalamus,therebysuppressingfoodintakeandstimulating energyexpenditure.Leptinreceptors–alternativelysplicedformsexistthatdiffer inlength–belongtothecytokineclassIreceptorfamily.Theyarefoundubiquitouslyin thebodyindicatingageneralroleofleptin,whichiscurrentlynotfullyunderstood.

Acirculatingformoftheleptinreceptorexistswhichactsasoneofseveralleptinbinding proteins.Besidesitsmetaboliceffects,leptinwasshowntohaveastronginfluenceona numberofendocrineaxes.Inmalemice,itbluntedthestarvation-inducedmarkeddeclineofLH, testosterone,thyroxineandtheincreaseofACTHandcorticosterone.Infemalemice,leptin preventedthestarvation-induceddelayinovulation.Ob/obmice,whichareleptindeficient duetoanobgenemutation,areinfertile.ThisdefectcouldbecorrectedbyadmiNISTrationof leptin,butnotthroughweightlossduetofasting,suggestingthatleptinispivotalfor reproductivefunctions.

Alltheseactionsmay,atleastinpart,beexplainedbythesuppressiveeffectofleptinon neuropeptideY(NPY)expressionandsecretionbyneuronsinthearcuatenucleus. NPYisastrongstimulatorofappetiteandisknowntobeinvolvedintheregulationof variouspituitaryhormones,e.g.suppressionofGHthroughstimulationofsomatostatin, suppressionofgonadotropinsorstimulationofthepituitary-adrenalaxis.

Themostimportantvariablethatdeterminescirculatingleptinlevelsisbodyfatmass. Obviously,underconditionsofregulareatingcycles,leptinreflectstheproportionofadipose tissueshowinganexponentialrelationship.Thisconstitutivesynthesisofleptinis modulatedbyanumberofnon-hormonalandhormonalvariables.Stimulatorsinbothrodents andhumansareoverfeeding,insulinandglucocorticoids. Suppressionhasbeenshownforfasting,cAMPandbeta-3-adrenoceptoragonists. Fromthesefindingsitbecomesclearthatleptinisanintegralcomponentof researchwithin metabolic andendocrinefeedbackloops.

 

品牌介绍

Mission Statement

Diapharma Group, Inc. in West Chester, Ohio sells hemostasis, thrombosis, platelet function testing, instrumentation, and apoptosis products in the diagnostic and research fields and provides strong technical competence and experience to ensure customer expectations will be met or exceeded.

diapharma hemostasis chromogenic clotting elisa assay test kit

 

History

The Diapharma Group, Inc. formed on January 1, 1997 from Pharmacia Hepar, Inc. in Franklin, Ohio, and began as the exclusive US and Canadian distributor of the Chromogenix substrates and assays.

Over a quarter-century ago, Chromogenix developed the first chromogenic substrate technology under the former name, Kabi Diagnostica. Kabi later merged with Pharmacia. Some of Diapharma’s current staff worked inside the Chromogenix department within Pharmacia’s heparin manufacturing plant.

In 1998, Diapharma moved to West Chester, Ohio, where it remains today. Over the years, Diapharma expanded its product line to include a variety of hemostasis, cell death, platelet function, ecotoxicology, assays, reagents, antibodies and instruments from superior manufacturers.

In 2017, Diapharma celebrated twenty years of success


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