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Product Details:
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Specifications: | 96 Wells/kit | Sensitivity: | 0.5ppb |
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Shelf Life: | 12 Months When Store At 2~8℃ | Detection Limit: | 2ppb |
Recovery Rate: | 70%~120% | Incubation Time: | 30min—15min |
Incubation Temperature: | 25℃ | ||
High Light: | veterinary elisa diagnostic kit,Enrofloxacin test kit,drug residue screening |
Enrofloxacin ELISA Kit
Catalog No. LSY-10017
1. Principle
This test kit is based on the indirect competitive enzyme immunoassay for the detection of Enrofloxacin in samples. The coupling antigens are pre-coated on the micro-well stripes. The Enrofloxacin in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti- Enrofloxacin antibodies. After the addition of the enzyme conjugate, the TMB substrate is added for coloration. The optical density (OD) value of the sample has a negative correlation with the Enrofloxacin in the sample. This value is compared to the standard curve and the Enrofloxacin concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 0.5 ppb
Incubation Temperature: 25℃
Incubation Time: 30min—15min
Detection limit
Tissue (shrimp)................................................................................. 2 ppb
Cross-reaction rate:
Enrofloxacin..................................................................................... 100%
Recovery rate:
Tissue (shrimp)....................................................................... 70%~120%
3. Components
1 | Micro-well strips | 12 strips with 8 removable wells each | |
2 | 6× standard solution (1 mL each) | 0ppb | 0.5ppb |
1.5ppb | 4.5ppb | ||
13.5ppb | 40.5ppb | ||
3 | High concentration standard | 1ml | 1 ppm |
4 | Enzyme conjugate | 7ml | red cap |
5 | Antibody working solution | 7ml | blue cap |
6 | Substrate A solution | 7ml | white cap |
7 | Substrate B solution | 7ml | black cap |
8 | Stop solution | 7ml | yellow cap |
9 | 20× concentrated washing buffer | 15ml | white cap |
10 | 2X Extracting buffer | 50ml*2 | transparent cap |
11 | 20× concentrated redissolving solution | 10ml | black cap |
4. Materials required but not provided
5. Sample pre-treatment
Instructions (The following points must be dealt with before the pre-treatment)
Solution preparation before sample pre-treatment:
5.1 Sample preparation of Tissue(shrimp)
1) Weigh 2.0 ± 0.05 g of the homogenized tissue sample into 50 ml centrifuge tube, add 2mL Extracting buffer and 8mL Ethyl acetate, shake for 3min, centrifuge at above 4000 r/min at room temperature for 5 min,
2) Take 1 ml up-layer liquid into a new 10mL centrifuge tube, blow to dry with nitrogen or air completely at 50 ℃
Fold of dilution of the sample: 4
6. ELISA procedures
6.1 Instructions
6.2 Operation procedures
7. Result judgment
There are two methods to judge the results; the first one is the rough judgment, while the second is the quantitative determination. Note that the OD value of the sample has a negative correlation with the content of Enrofloxacin in the sample.
7.1 Qualitative determination
The concentration range (ng/mL) obtained from the comparison the average OD value of the sample with that of the standard solution. Assuming that the OD value of the sampleⅠ is 0.3, and that of the sampleⅡ is 1.0, the OD value of standard solutions is: 2.243 for 0ppb, 1.816 for 0.5ppb, 1.415 for 1.5ppb, 1.8 for 4.5ppb, 0.313 for 13.5ppb ,0.155 for 40.5ppb, accordingly the concentration range of the sampleⅠ is 13.5 to 40.5ppb, and that of the sampleⅡ is 1.5 to 4.5ppb. (multiplied by the corresponding dilution fold)
7.2 Quantitative determination
The mean values of the absorbance values is equivalent to the percentage of the average OD value (B) of the sample and the standard solution divided by the OD value (B0) of the first standard solution (0 standard) and subsequently multiplied by 100%, that is,
Percentage of absorbance value = | B | ×100% |
B0 |
B—the average (double wells) OD value of the sample or the standard solution
B0—the average OD value of the 0ng/mL standard solution
Draw the standard curve with the absorption percentages of the standard solutions and the semilogarithm values of the Enrofloxacin standard solutions (ng/mL) as Y- and X-axis, respectively. Read the corresponding concentration of the sample from the standard curve by incorporating its absorption percentage into the standard curve. The resulting value is subsequently multiplied by the corresponding dilution fold, finally obtaining the Enrofloxacin concentration in the sample.
Using the professional analyzing software of this kit will be more convenient for the accurate and rapid analysis of a large amount of samples. (Please contact us for this software)
8. Precautions
8. The optimum reaction temperature is 25 ℃, and too high or too low temperatures will result in the changes in the detecting sensitivity and OD values.
9. Storage and expiry date
Storage: store at 2-8 ℃, not frozen.
Expiry date: 1 year; date of production is on box.
Note: If the Vacuum package of microplate has leakage, it is still valid to use, do not affect the test result, be relax to use.