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Salbutamol ELISA Kit

Salbutamol ELISA Kit

Hersteller	Creative Diagnostics
Kategorie	Food Safety Drugs ELISA ELISA & Immunoassays ELISA Kits Immunoassay
Typ	Elisa-Kit
Specific against	other
Menge	96T
ArtNr	DEIA027
eClass 6.1	32160605
eClass 9.0	32160605
Lieferbar
Abbr	Salbutamol Kit
Storage	Unopened Kit: Store at2 - 8oC. Do not use past kit expiration date. Opened/ReconstitutedReagents: TMB Solution A, TMB Solution B, TMB Stop Solution, WashBuffer, HRP-conjugate antibody The above mentionedreagents should be stored for up to 1 month at 2 - 8oC. Microplate Wells: Returnunused wells to the foil pouch containing the desiccant pack, reseal alongentire edge of zip-seal. May be stored for up to 1 month at 2 - 8oC.
Principle Of The Test	The method is basedon an indirect competitive ELISA assay. The drug of interest has been coatedin the plate wells. During the analysis, sample is added along with theprimary antibody specific for the target drug. If the target is present inthe sample, it will compete for the antibody, thereby preventing the antibodyfrom binding to the drug attached to the well. The secondary antibody, taggedwith a peroxidase enzyme, targets the primary antibody that is complexed tothe drug coated on the plate wells. The resulting color intensity, afteraddition of substrate, has an inverse relationship with the targetconcentration in the sample.
Reagents And Materials Provided	Microplate:96well polystyrene microplate (12 strips of 8 wells) coated with Salbutamol, Salbutamol Standards(6): 0, 0.3, 0.9, 2.7, 8.1, 24.3ppb, 6vials, Highconcentration Salbutamol Standards: 1ppb, 2ml, 1 vial, Antibody Solution: 7ml, 1 vial, HRPConjugate Antibody: 12ml, 1 vial, Sample Diluent(10×): 30 ml. Use to dilutesamples. Wash Solution (20×) Concentrate: 40 ml, 1vial, TMB Solution A: 7ml, 1vial, TMB Solution B: 7ml, 1vial, TMB Stop Solution: 7ml, 1vial, Microtiter plate sealers Plastic Sealable Bag
Materials Required But Not Supplied	Equipment: 1. Validatedmicroplate reader. 2. Homogenizer 3. Electronicbalance 4. Centrifuger 5. Shaker formicrotiter plates (optional) 6. Organomation 7. Vortex genie 8. Validatedadjustable micropipettes, single and multi-channel. 9. Timer
Precautions	1. The kit should beequilibrated to room temperature (20-23C) before opening any vials and starting the assay. Itis highly recommended that the solutions be used as soon as possible afterrehydration. 2. Do not mix orsubstitute reagents with those from other lots or sources. 3. To avoidcross-contamination, change pipette tips between additions of each standardlevel, between sample additions, and between reagent additions. Also, useseparate reservoirs for each reagent. 5. Crystals couldappear in the 20X wash solution due to high salt concentration in the stocksolutions. Crystals are readily dissolved at room temperature or at 37oCbefore dilution of the buffer solutions. 6. Keep TMBSubstrate protected from light. 7. The Stop Solutionprovided with this kit is an acid solution. Wear eye, hand, face, andclothing protection when using this material.
Specimen Treatment	Homogenized, centrifuged, evaporated, diluted for assay.
Detection Method	Competitive Enzyme Immunoassay
Sample	feed, urine and Tissue (pork?pork liver)
Indications Of Instability Or Deterioration Of The Reagents	1. Values of thePositive or Negative controls, which are out of the indicated Quality controlrange, are indicator of possible deterioration of the reagents and/oroperator or equipment errors. In such case, the results should be consideredas invalid and the samples must be retested. In case of constant erroneousresults classified as due to deterioration or instability of the reagents, immediately substitute the reagents with new ones, or contact our technicalsupport for further assistance. 2. If after mixingof the TMB Solution A and B into the wells, the color of the mixtureturns blue within few minutes, do not continue carrying out the testing andreplace the reagents with fresh ones.
Assay Procedure	1. Add 50ul of thestandard solutions or samples (sample extracts) into the wells of the teststrips according to the working scheme given. We recommend using duplicatesor triplicates. 2. Add 50ul of antibodysolution to the individual wells successively using a multi-channel pipetteor a stepping pipette. 3. Cover the wellswith parafilm or tape and mix the contents by moving the strip holder in acircular motion on the benchtop for about 30 seconds. Be careful not to spillcontents. 4. Incubate thestrips for 40 minutes at room temperature. 5. After incubation, remove the covering and vigorously shake the contents of these wells into asink. Wash the strips three times using the 1X washing buffer solution. Useat least a volume of 260 ul of washing buffer for each well and each washingstep. Remaining buffer in the wells should be removed by patting the platedry on a stack of paper towels. 6. Dispense 50 ul ofTMB Solution A and 50 ul TMB Solution B into each well. Cover the wells withparafilm or tape and mix the contents by moving the strip holder in acircular motion on the benchtop for about 30 seconds. Incubate the strips for15-20 minutes at room temperature. Protect the strips from direct sunlight. 7. Add 50 ul of stopsolution to the wells in the same sequence as for the substrate solution. 8. Read theabsorbance at 450 nm and 630 nm using a microplate ELISA photometer within 5minutes after the addition of the stopping solution.
Evaluation	The evaluation ofthe ELISA can be performed using commercial ELISA evaluation programs(4-Parameter (preferred) or Logit/Log. For manual evaluation, calculate themean absorbance value for each of the standards. Calculate the %B/B0 for eachstandard by dividing the mean absorbance value for each standard by the ZeroStandard (Standard 0) mean absorbance. Construct a standard curve by plottingthe %B/B0 for each standard on the vertical linear (y) axis versus thecorresponding Salbutamol concentration on the horizontal logarithmic (x) axison graph paper. %B/B0 for samples will then yield levels in ppb of a Salbutamolby interpolation using the standard curve. Samples showing lowerconcentrations of Salbutamol compared to Standard 1 (0.3 ng/mL) areconsidered as negative. Samples showing a higher concentration than Standard5 (24.3 ng/mL) must be diluted further to obtain accurate results.

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