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REAGEN™ patulin ELISA Test Kit enables government agencies, food manufacturers and quality assurance organizations to detect patulin in various sample types and to satisfy customer concerns about food safety.
1.The extraction of patulin from feed, honey, meat, milk, tissue, serum and urine can be finished in 10 - 30 minutes with a high recovery rate of 75 - 95%.
2. A quick ELISA assay (less than 2 hours regardless of number of samples).
3. High reproducibility.
The method is based on a competitive colorimetric ELISA assay. The patulin antibody has been coated in the plate wells. During the analysis, sample is added along with the patulin-horseradish peroxidase (patulin-HRP) conjugate. If the patulin residue is present in the sample, it will compete for the patulin antibody, thereby preventing the patulin-HRP from binding to the antibody attached to the well. The resulting color intensity, after addition of the HRP substrate (TMB), has an inverse relationship with the patulin residue concentration in the sample.
REAGEN™ patulin 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.
1.The standards contain patulin. Handle with particular care.
2.Do not use the kit past the expiration date.
3.Do not mix reagents from different kits or lots except for components with the same part No’s within their expiration dates. HRP CONJUGATES AND PLATES ARE KIT-AND LOT-SPECIFIC
4.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 insulation material under the assay plates during incubation.
5.Make sure you are using only distilled or deionized water since water quality is very important.
6.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.
7.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.
8.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.
9.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.
| Sample Type | Detection Limit (ng/g or ppb) |
| Feed | 1 |
| Honey | 2 |
| Meat/Liver/Kidney/Fish/Shrimp | 1 |
| Milk | 0.5 |
| Serum | 0.5 |
| Urine | 0.5 |
| Juice | 1 |
| Name | Cross-Reactivity (%) |
| patulin | 100 |
| Oxolinic acid | 40 |
| Pipemidic acid | 18 |
| Ofloxacin | 17 |
| Ciprofloxacin | 9 |
| Enrofloxacin | 6 |
| Flumequin | 6 |
| Cinoxacin | 1 |
1. Microtiter plate reader (450 nm)
2. Incubator
3. Tissue Mixer (e.g. Omni TissueMaster Homogenizer)
4. Vortex mixer (e.g. Gneie Vortex mixer from VWR)
5. 10, 20, 100 and 1000 µL pipettes
6. Multi-channel pipette: 50-300 µL (Optional)
1.The standards contain patulin. Handle with particular care.
2.Do not use the kit past the expiration date.
3.Do not mix reagents from different kits or lots except for components with the same part No’s within their expiration dates. HRP CONJUGATES AND PLATES ARE KIT-AND LOT-SPECIFIC
4.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 insulation material under the assay plates during incubation.
5.Make sure you are using only distilled or deionized water since water quality is very important.
6.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.
7.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.
8.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.
9.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.
To a reasonable amount of sample, add 5 times the amount of 70% methanol (e.g. 1 g of sample + 5 mL of 70% methanol); homogenize the sample with a suitable mixer.
Take out 1.5 mL of the homogenized sample and centrifuge for 5 minutes at 4,000 x g at room temperature (20 – 25°C / 68 – 77°F).
Transfer 0.5 mL of the supernatant to a tube, add 0.5 mL of 1X Sample Extraction Buffer, and mix well.
Use 100 µL of the sample per well for the assay.
Note: Dilution factor: 10.
Take out 0.1 g of honey sample, add 1.9 mL of 35% Methanol/Sample Extraction Buffer and mix well.
Use 100 µL per well for the assay.
Note: Dilution factor: 20
Remove fat from the sample. Homogenize the sample with a suitable mixer.
Weigh out 1.0 g of the homogenized sample and add 4 mL of 70% methanol.
Vortex for 10 minutes at maximum speed.
Centrifuge for 5 minutes at 4,000 x g at room temperature (20 – 25°C / 68 – 77°F).
Transfer 0.5 mL of the supernatant to a tube, add 0.5 mL of 1X Sample Extraction Buffer, and mix well.
Use 100 µL for the assay.
Note: Dilution factor: 10.
For fat-free milk, take out 200 µL of the milk sample, add 800 µL of 35% Methanol/Sample Extraction Buffer, and mix well. Take 100 µL of the diluted sample per well for the assay.
For regular milk with fat, centrifuge 1 mL of the milk sample at 4,000 x g for 5 minutes, discard the upper fat layer. Take out 200 µL of the milk sample, add 800 µL of 35% Methanol/Sample Extraction Buffer, and mix well. Take 100 µL of the diluted sample per well for the assay.
Note: Dilution factor: 5.
Centrifuge 0.5 mL of serum at 4,000 x g for 5 minutes.
Take out 0.2 mL of the supernatant, add 0.8 mL of 35% Methanol/Sample Extraction Buffer.
Use 100µL per well for the assay.
Note: Dilution factor: 5.
