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Topic Melamine ELISA Test Kit 
Name NICOLE 
Company REAGEN 
Address 102 Commerce Dr. Unit 8, Moorestown, NJ 08057  
Phone  
Mobile  
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Country USA 
Date Created 9/28/2013 11:58:11 AM 
Message Product Description
REAGEN™Melamine ELISA Test Kit is a competitive enzyme immunoassay for the quantitative analysis of melamine in milk, milk powder, infant formula, meat, fish, feed, chocolate and other food samples. The unique features of the kit are:
 High recovery (80-105%),rapid(10-40 minutes),and cost-effective extraction methods.
 High sensitivity(5ng/g or ppb)and low detection limit for liquid milk (0.2ppm).
 High reproducibility.
 A quick ELISA assay(less than 2 hours regardless of number of samples).

Procedure Overview
The method is based on a competitive colorimetric ELISA assay. The drug of interest has been coated in the plate wells. During the analysis, sample is added along with the primary antibody specific for the target toxin and enzyme-conjugate. If the target is present in the sample, it will compete for the antibody, thereby preventing the antibody from binding to the drug attached to the well. The enzyme-conjugate, will bind with the primary antibody that is complexed to the drug coated on the plate wells. The resulting color intensity, after addition of substrate, has an inverse relationship with the target concentration in the sample.

Kit Contents, Storage and Shelf Life
REAGEN™Melamine ELISA Test Kit has the capacity for 96 determinations or testing 、of 42 samples in duplicate (assuming 12 wells for standards). Return any unused microwells to the foil bag and reseal them with the desiccant provided in the original package. Store the kit at 2-8°C *. The shelf life is 12 months when the kit is properly stored.
Kit Contents Amount Storage
Melamine Coated Plate 1 x 96-well Plate
(8 wells x 12 strips) 2-8°C
Melamine Standards:
Negative control (white cap tube)
5 ng/mL (yellow cap tube)
20 ng/mL (orange cap tube)
50 ng/mL (pink cap tube)
100 ng/mL (purple cap tube)
200 ng/mL (blue cap tube)
100 µg/mL (spiking, optional, red cap tube)
1.0 mL
1.0 mL
1.0 mL
1.0 mL
1.0 mL
1.0 mL
1.0 mL




2-8°C *
Melamine Antibody 8 mL 2-8°C
100X HRP Conjugate 100 µL
HRP Conjugate Diluent 10 mL
20X Wash Solution 30 mL
Stop Buffer 20 mL
TMB Substrate 10 mL
10 X Melamine Sample Extraction Buffer
1 X Melamine Clean Up Buffer
1X Melamine Sample Balance Buffer
10X Melamine Sample Dilution Buffer 20 mL X 2
20 mL
20 mL
20 mL

*If you are not planning to use the kit for over 1 month, store Melamine Standards, Melamine Antibody and 100X HRP Conjugate at -20°C or in a freezer.

Sensitivity (Detection Limit)

Sample Type Detection Limit (g/g or ppb)
Feed 0.5 ppm
Meat/Fish 0.2 ppm
Milk 0.2 ppm
Milk Powder/Infant Formula 0.4 ppm
Chocolate 0.4 ppm

Specificity (Cross-Reactivity)

Analytes Cross-Reactivity (%)
Melamine 100
Cyanurate 15
Ammeline 5
Ammelide <1

Required Materials Not Provided With the Kit

 Microtiter plate reader (450 nm)
 Tissue Mixer (e.g. Omni TissueMaster Homogenizer)
 Vortex mixer (e.g. Genie Vortex mixer from VWR)
 10, 20, 100 and 1000 µL pipettes
 Multi-channel pipette: 50-300 µL (Optional)

Warnings and Precautions

 The standards contain melamine. Handle with particular care.
 Do not use the kit past the expiration date.
 Do not intermix reagents from different kits or lots except for components with the same part No’s within their expiration dates. ANTIBODIES AND PLATES ARE KIT-AND LOT-SPECIFIC. Make sure that the HRP Conjugate and Diluent are mixed in correct volumes.
 Try to maintain a laboratory temperature of 20°–25°C (68°–77°F). Avoid running assays under or near air vents, as this may cause excessive cooling, heating and/or evaporation. Also, do not run assays in direct sunlight, as this may cause excessive heat and evaporation. Cold bench tops should be avoided by placing several layers of paper towel or some other insulate material under the assay plates during incubation.
 Make sure you are using only distilled or deionized water since water quality is very
important.
 When pipetting samples or reagents into an empty microtiter plate, place the pipette tips in the lower corner of the well, making contact with the plastic.
 Incubations of assay plates should be timed as precisely as possible. Be consistent when adding standards to the assay plate. Add your standards first and then your samples.
 Add standards to plate only in the order from low concentration to high concentration, as this will minimize the risk of compromising the standard curve.
Always refrigerate plates in sealed bags with a desiccant to maintain stability. Prevent condensation from forming on plates by allowing them equilibrate to room temperature (20 – 25°C /68 – 77°F) while in the packaging.
 
 
      
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