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Topic Colistin ELISA Test Kit 
Name NICOLE 
Company REAGEN 
Address 102 Commerce Dr. Unit 8, Moorestown, NJ 08057  
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Country USA 
Date Created 9/27/2013 2:52:14 PM 
Message Product Description
REAGEN™ Colistin ELISA Test Kit is a competitive enzyme immunoassay for the quantitative analysis of Colistin in meat (beef, pork and chicken) and Egg.Colistin, also known as polymyxin E, is a cationic lipopeptide antibiotic isolated in 1950 from Bacillus polymyxa subsp. colistinus. It is most notable for its activity toward the multidrug-resistant organism Pseudomonas aeruginosa and low rates of resistance. Colistin is a mixture of cyclic polypeptides colistin A and B. Colistin is effective against most Gram-negative bacilli. The REAGEN™ Colistin ELISA Test Kit enables international and government regulatory agencies, food manufacturers and processors, as well as quality assurance organizations, to detect colistin in various sample types and to satisfy customer concerns about food safety
The unique features of the kit are:
 High recovery, rapid, and cost-effective extraction methods
 High sensitivity (0.5 ng/g or ppb)
 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 drug. 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 secondary antibody, tagged with a peroxidase enzyme, targets 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™ Colistin 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
Colistin-coated Microtiter Plate 1 x 96-well plate (8 wells x 12 strips) 2-8C
Standards:
Negative control
0.5 ng/mL
1.5 ng/mL
5 ng/mL
15 ng/mL
50 ng/mL
1000 ng/mL (spiking, optional)
0.8mL
0.8mL
0.8mL
0.8mL
0.8mL
0.8mL
0.8mL 2-8C *
Colistin Antibody #1(powder) 3 tubes 2-8C
Antibody #1 Diluent 14 mL
100X HRP-Conjugated Antibody #2 250 L
Antibody #2 Diluent 20 mL
10X Colistin Extraction Buffer 25 mL
20X Wash Solution 28mL
Stop Buffer 14 mL
TMB Substrate 10 mL
Clean Up Buffer I (Optional)
Clean Up Buffer II (Optional) 18 mL
18mL
* If you are not planning to use the kit for over 1 month, store Colistin Standard , Colistin Antibody #1, 100X HRP-Conjugated Antibody #2 at -20C preferably or in a freezer.
Sensitivity

Sample Type Detection Limit (ng/g or ppb)
Meat(Beef/Pork/Chicken) 4
Egg 50

Specificity

Molecules Cross-Reactivity (%)
Colistin 100
Polymyxin 88
Bacitracin zinc < 0.5

Required Materials Not Provided With the Kit

 Microtiter plate reader (450 nm)
 Incubator
 Tissue Mixer (e.g. Omni TissueMaster Homogenizer)
 Vortex mixer (e.g. Gneie Vortex mixer from VWR)
 20, 100 and 1000 L pipettes
 Multi-channel pipette: 50-300 L (Optional)
 Acetonitrile,hexane
 
 
      
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