REAGEN™Beta-Lactam ELISA Test Kit is a competitive enzyme immunoassay for the quantitative analysis of beta-lactams including ampicillin, penicillin G, amoxicillin, cloxacillin, oxacillin in milk, urine, tissue (liver, kidney, and muscle), serum, and plasma samples.REAGEN™Beta-lactam ELISA Test Kit enables international and government regulatory agencies, food manufacturers and processors, as well as quality assurance organizations, to detect beta-lactam containing antibiotics to satisfy customer concerns about food safety.
The unique features of the kit are:
§ High recovery (>80%), low-cost rapid sample prep protocol (10 minutes - 1 hour)
§ High sensitivity: 0.4 ng/gram = 0.4 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%)
The method is based on a competitive colorimetric ELISA assay. The drug of interest (beta-lactam antibiotic) has been coated in the plate wells. The sample to be tested is mixed with a high-affinity beta-lactam capture protein and added to the well. If the beta-lactam 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 beta-lactam antibiotic in the sample are determined by comparison of the color intensity of unknown samples with the standard curve, which is made using the beta-lactam standard provided in the kit.
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Kit Contents
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Amount
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Storage
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Beta-Lactam Coated Plate
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1 x 96-well Plate (8 wells x 12 strips)
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2-8°C
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Penicillin G powder stock (1000 ng):
Empty Vials for Standards:
Negative control (white cap tube)
0.04 ng/mL (yellow cap tube)
0.08 ng/mL (orange cap tube)
0.2 ng/mL (pink cap tube)
0.3 ng/mL(purple cap tube)
0.6 ng/mL (blue cap tube)
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2
1
1
1
1
1
1
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2-8°C *
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10X Beta-Lactam Capture Protein
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1.8 mL
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2-8°C *
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Milk Standard Dilution Buffer 1
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15 mL
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2-8°C *
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200X HRP-Conjugated Antibody #2
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100 mL
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2-8°C *
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Antibody #2 Diluent **
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20 mL
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2-8°C
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10X Beta-Lactam Extraction Buffer*
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2 x 28 mL
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2-8°C
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20X Wash Solution **
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28 mL
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2-8°C
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Stop Buffer **
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20 mL
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2-8°C
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TMB Substrate **
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12 mL
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2-8°C
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* If the kit is to be stored for over two weeks before use, store Standard Dilution Buffer1, Penicillin G powder stock, 10X Beta-Lactam Capture Protein and 100X 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
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Antibiotic
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Milk, Meat, Tissue, Fish and Urine(ppb)
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Serum and Plasma(ppb)
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Penicillin G
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0.40
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0.80
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Ampicillin
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0.43
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0.86
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Azlocillin
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0.40*
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0.80
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Pipercillin
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0.52
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1.04
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Nafcillin
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0.58
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1.16
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Amoxicillin
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0.62
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1.24
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Cloxacillin
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0.64
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1.28
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Oxacillin
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0.64
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1.28
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Cefoperazone
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0.67
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1.34
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Ceftiofur
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0.7
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1.4
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