Product Description
REAGEN Ceftiofur ELISA Test Kit is a competitive enzyme immunoassay for the quantitative analysis of Ceftiofur in milk, urine, tissue (liver, kidney, and muscle), serum, embryonic eggs, and plasma samples.
The unique features of the kit are:
- High recovery (>80%), low-cost rapid sample prep protocol (10 minutes – 1 hour)
- High sensitivity: 2 ng/gram = 2 ppb (parts per billion)
- Rapid assay protocol (less than 2 hours to process 40 samples in duplicate)
- High reproducibility (interplate and intraplate CV typically <8%)
Procedure Overview
REAGEN The method is based on a competitive colorimetric ELISA assay. The drug of interest (Ceftiofur) has been coated in the plate wells. The sample to be tested is mixed with a high-affinity Ceftiofur capture protein and added to the well. If the Ceftiofur target is present in the sample, it will compete for the capture protein, thereby preventing the capture protein from binding to the drug attached to the well. The secondary antibody, tagged with a peroxidase enzyme, targets the capture protein bound 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. The levels of Ceftiofur in the sample are determined by comparison of the color intensity of unknown samples with the standard curve, which is made using the Ceftiofur standard provided in the kit.
Kit Contents, Storage and Shelf Life
REAGEN Ceftiofur 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 |
| Ceftiofur Coated Plate | 1 x 96-well Plate (8 wells x 12 strips) | 2-8°C |
| Ceftiofur powder stock (1000 ng):
Empty Vials for Standards: Negative control (white cap tube) 2 ng/mL (yellow cap tube) 8 ng/mL (orange cap tube) 16 ng/mL (pink cap tube) 32 ng/mL (purple cap tube) 64 ng/mL (blue cap tube) |
2
1 1 1 1 1 1 |
2-8°C * |
| 10X Ceftiofur Capture Protein | 1.8 mL | 2-8°C * |
| Capture Protein Diluent | 12 mL | 2-8°C * |
| Milk Standard Dilution Buffer 1 (Optional) | 15 mL | 2-8°C * |
| 200X HRP-Conjugated Antibody #2 | 100 mL | 2-8°C * |
| Antibody #2 Diluent ** | 20 mL | 2-8°C |
| 10X Sample Extraction Buffer** | 2 x 25 mL | 2-8°C |
| 20X Wash Solution ** | 28 mL | 2-8°C |
| Stop Buffer ** | 14 mL | 2-8°C |
| TMB Substrate ** | 12 mL | 2-8°C |
* If the kit is to be stored for over two weeks before use, store Standard Dilution Buffer1, Ceftiofur powder stock, 10X Ceftiofur Capture Protein and 200X HRP-Conjugated Antibody #2 at -20°C in a freezer that does not cycle to higher temperatures (i.e. a non-frost-free freezer). To avoid repeated freeze-thawing of the capture protein it is strongly recommended to store this component in single-use aliquots.
Sensitivity (Detection Limit)
| Sample Type | Detection Limit (ng/g or ppb) |
| Milk, Meat, Tissue, Fish and Urine, Embryonic Eggs | 20 |
| Serum and Plasma | 40 |
Specificity (Cross-Reactivity)
| Ceftiofur | Cross-Reactivity (%) |
| Ceftiofur | 100 |
| Cefoperazone | 102 |
| Oxacillin | 193 |
| Cloxacillin | 195 |
| Amoxicillin | 208 |
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