Veratox® for Chloramphenicol

Item No.  9551

  • Rapid results easily determined by reader
  • Low levels of detection for Minimum Required Performance Limit (MRPL)
$581.00
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Veratox® for Chloramphenicol is used for the quantitative analysis of chloramphenicol residues in shrimp. The test is a competitive direct ELISA that provides exact concentrations in parts per trillion (ppt). Free chloramphenicol in the sample and controls competes with enzyme-labeled chloramphenicol (conjugate) for the antibody binding sites. After a wash step, substrate reacts with the bound enzyme conjugate to produce blue color. A microwell reader is used to yield optical densities. Control optical densities form a standard curve, and sample optical densities are plotted against the curve to calculate the exact concentration of chloramphenicol.
Specifications
Analyte Chloramphenicol
Brand Veratox®
Limit Of Detection 0.02 ng/mL or 10 ppt
Package Dimensions 6.10 IN x 3.10 IN x 4.10 IN
Package Weight 1.50 LB
Quantity per Package 90 tests

  1. 1 plate (96 wells) of antibody-coated microwells
  2. 1 vial of chloramphenicol 0.0 ng/mL standard, 2 mL
  3. 1 vial of chloramphenicol 0.02 ng/mL standard, 2 mL
  4. 1 vial of chloramphenicol 0.05 ng/mL standard, 2 mL
  5. 1 vial of chloramphenicol 0.1 ng/mL standard, 2 mL
  6. 1 vial of chloramphenicol 0.5 ng/mL standard, 2 mL
  7. 1 vial of chloramphenicol 2 ng/mL standard, 2 mL
  8. 1 bottle of ready-to-use chloramphenicol enzyme conjugate, 20 mL
  9. 1 bottle of sample dilution buffer, 100 mL
  10. 1 bottle of wash buffer concentrate, 100 mL (dilute 1:10 with deionized water before use)
  11. 1 bottle of substrate solution, 28 mL
  12. 1 bottle of stop solution, 28 mL
  1. Pipettor, 100 µL (Neogen® item #9272)
  2. Pipettor, multichannel, 100 µL
  3. Pipette tips for 100 µL and multichannel pipettors (Neogen item #9410)
  4. 3 reagent boats for multichannel pipettor
  5. Paper towels or equivalent absorbent material
  6. Pipettor capable of measuring 1.0 mL
  7. Pipettor capable of measuring 0.5 mL (for isooctane/chloroform)
  8. Graduated or volumetric pipettor capable of measuring 6.0 mL of solvent
  9. Disposable pipettes capable of measuring 4.0 mL of solvent
  10. Glass test tubes capable of being centrifuged at 2,000 g (16 x 100 mm preferred)
  11. Caps for glass test tubes
  12. Ethyl Acetate (ACS reagent grade or better)
  13. Chloroform
  14. Isooctane (2,2,4-trimethylpentane)
  15. Deionized or distilled water
  16. Blender or food processor
  17. Centrifuge capable of 2,000 g, swinging bucket rotor preferred
  18. Heating block
  19. Compressed air or nitrogen and necessary tubing
  20. Water bath
  21. Vortex
  22. Automated plate washer
  23. Microplate reader with 450 nm filter (650 nm reference filter is optional)
  24. Balance capable of measuring 3.0 g
  25. Timer (Neogen item #9426)
  26. Graduated cylinder to dilute wash buffer
  27. Waterproof marker
  28. Pipette aid for dispensing solvents
  29. Stainless steel spatulas for transferring shrimp into test tubes
  30. Thermometer

Veratox for Chloramphenicol is an enzyme-linked immunosorbent assay (ELISA). The basis of this test is a competition between a chloramphenicol-enzyme conjugate and any free chloramphenicol from the sample for a limited number of binding sites on the antibody-coated microplate wells.

Samples are extracted, and the extracts (and solutions of chloramphenicol standards) are added to the microplate wells. A solution of the chloramphenicol-enzyme conjugate is also added to each of the wells. At this point, time is allowed for any free chloramphenicol and the chloramphenicol-enzyme conjugate to compete for the binding sites on the plate. Next, the plate is washed to remove unbound material and a substrate solution is added to each well. During an ensuing reaction period, any bound enzyme conjugate reacts with the substrate to produce a blue-coloured solution. The colour-forming reaction is terminated by adding Stop Solution to the wells of the plate, turning the blue colour to yellow. The absorbance of each well is then measured using a microplate reader set at 450 nm. The extent of colour formation is inversely proportional to the amount of chloramphenicol present in that well.

Samples that contain little or no chloramphenicol will have a relatively large absorbance while a sample that contains more chloramphenicol will have a smaller absorbance. Quantitative results may be obtained by comparing the absorbance of the samples to a standard curve generated from the wells containing the chloramphenicol standards.

Training

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