Alert® for Ractopamine is meant for the qualitative determination of trace quantities of ractopamine in tissue. Neogen’s® enzyme-linked immunosorbent assay (ELISA) products are for screening purposes only. It is recommended that all suspect samples are confirmed by a quantitative method, such as mass spectrometry.
Specifications
Analyte Ractopamine
Brand Alert®
Cross Reactivity Ractopamine 100%, Ritodrine 19.9%, Fenoterol 0.5%
Cut-Off 0.2 ppb cut-off level for muscle
Limit of Detection 0.2 ppb
Package Dimensions 6.50 IN x 4.30 IN x 4.60 IN
Package Weight 0.50 LB
Platform ELISA
Prop 65 Warning
Warning
WARNING: This product can expose you to chemicals including thimerosal, which is known in the State of California to cause birth defects or other reproductive harm. For more information, go to www.P65Warnings.ca.gov.
Quantity per Package 92 tests
Result Type Qualitative
Testing Time 90.00 - 120.00 Minutes
Time to Result 20 Minutes

  1. Antibody-coated strips in well holder 1 x 96-well plate (8 wells x 12 strips)
  2. Ractopamine Standards:
    1. 0 ng/mL Muscle Standard 0.5 mL
    2. 0.2 ng/mL Muscle Standard 0.5 mL
  3. Ractopamine Enzyme 1 vial, lyophilized
  4. Enzyme Diluent 16 mL
  5. Sample Diluent 125 mL
  6. 10X Wash Solution 60 mL
  7. K-Blue® Substrate 13 mL
  8. Stop Solution 13 mL

Reagents

  1. Ethyl acetate, HPLC grade (Fisher E195-4)
  2. Sodium carbonate, anhydrous (Fisher S263-1)
  3. Distilled water
  4. Sodium Acetate, Bioxtra = 99% (Sigma Cat# S-7545)
  5. Acetic acid, ACS reagent = 99.7% purity (Sigma Cat# 320099)
  6. β-glucuronidase Type HP-2 from Helix pomatia. (Sigma Cat# G-7017)

Materials

  1. Disposable culture tubes or comparable vessels (glass preferred)
  2. 5 mL or 10 mL serological pipettes (Neogen item 9415)
  3. Pipettors and tips capable of delivering 50 µL and 100 µL (Neogen items 9276, 9407, 9417)
  4. Well holder (Neogen item 9402) (NOTE: One well holder is included in kit, additional holders are necessary to run test.)
  5. Reagent troughs or weigh boats for dispensing reagents (Neogen items 9450 or 9435)
  6. Multichannel pipettor (NOTE: Not required if using automated liquid handling equipment) (Neogen items 9385, 9407, 9417)
  7. Vortex (Fisher item 02-215-365)
  8. Plate washer, multichannel pipettor or squirt bottle (Neogen items 9400, 9366)
  9. Microplate reader, 450 nm wavelength absorbance filter (650 nm, 630 nm, or 620 nm wavelength reference filter) (Neogen item 9303)
  10. 50 mL polypropylene disposable centrifuge tubes (Fisher 06-442-19)
  11. 16 x 100 mm borosilicate disposable glass culture tubes and caps (Fisher 14-961-29 and 14-376-84)
  12. Centrifuge and rotor with inserts (Fisher item 05-376-199 and 05-376-237)
  13. Electric pipette aid (Fisher item 13-681-15)
  14. Pipette pumps, 2-10 mL (Neogen item 9277)
  15. Pipette pump, 2 mL (Fisher item 13-683-1B)
  16. Thermometer (Neogen item 9461)
  17. Timers, multichannel (Neogen item 9426)
  18. Tube rack for 16 x 100 mm culture tubes, 20 positions (Fisher item 14-792-12)
  19. Tube rack for 50 mL centrifuge tubes, 16 positions (Fisher item 14-791-6B)
  20. Chemi-Scraper spatula (Fisher item 14-373)

Each test kit contains antibody-coated microwells with antibodies specific to the kit’s target substance.

First, samples and controls are added to their respective test wells. Next, an enzyme conjugate (target substance chemically linked with an enzyme) is added. The samples/controls and conjugate are mixed and transferred to antibody wells where they compete for the antibody binding sites. The more target substance in the sample, the less conjugate that binds in the wells.

After an incubation, the wells are washed to remove all unbound materials.

A substrate, which changes color in the presence of the conjugate, is added to the wells. During an incubation, yellow color develops in proportion to the amount of conjugate versus target substance in the wells. Wells are read on a microwell reader with a 450 nm filter. Optical densities are obtained. If the optical density of the sample is quarter than the control, the sample contains less target residue than the control. Lower optical densities indicate positive samples.

Alert_yellow-wells.png

Training

Our customers’ success is our shared success. Our experts are ready to train you and your team on our solutions, so you can rest easy knowing procedures are performed properly and yield accurate results. In addition, we provide certificates upon completion of training to provide you with the documentation needed for audit traceability.


Support

Validation

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