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Product Details:
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Specifications: | 96 Wells/kit | Sample Performance: | Urine, Tissue |
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Sensitivity: | 0.05ppb | Incubator Temperature: | 25℃ |
Incubator Time: | 30min~15min | Shelf Life: | 12 Months When Properly Stored |
MOQ: | 1 Kit | ||
High Light: | Ractopamine ELISA,Food Safety ELISA,ract assay kit |
Ractopamine ELISA Test Kit
Catalog No. LSY-10013
1. Principle
The test kit is based on the competitive enzyme immunoassay for the detection of Ractopamine in the sample. The conjugated antigens is pre-coated on the micro-well stripes. The Ractopamine in the sample and the coupling antigens pre-coated on the micro-well stripes compete for anti-Ractopamine 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 Ractopamine concentration in the sample. The value is compared to the standard curve and the Ractopamine concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 0.05 ppb
Incubator temperature: 25℃
Incubator time: 30min~15min
Detection limit
Urine 0.1ppb
Tissue……………………………………………………………………… 0.2 ppb
Cross-reactions
Ractopamine 100%
Dobutamine <1%
Salbutamol <0.1%
Clenbuterol <0.1%
Recovery rate
Urine, tissue 60~120%
3. Components
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:
Solution I 0.2M HCl:
Take 17.2mL HCl, add deionized water to 1L.
Solution II 1M NaOH:
Take 4g NaOH, add deionized water to 100mL.
Solution Ⅲ Sample extracting solution:
Dilute 2× concentrated extracting solution with deionized water at 1:1.
Samples preparation
5.1 Urine
Take 50 µL clear urine, directly detect it (If urine is muddy, must filter or centrifuge at 4000 r/min for 10 min, then take clear urine). Store at frozen environment if don’t use.
Fold of dilution of sample: 1
5.2 Tissue
Fold of dilution of sample: 4
6. ELISA procedures
6.1 Instructions
6.2 Operation procedures
6 Coloration: add 50 µL of the substrate A solution, 50 µL of the B solution into each well. Mix gently by shaking the plate manually, and incubate at 25 ℃ for 15 min at dark for coloration.
7 Determination: add 50 µL of the stop solution into each well. Mix gently by shaking the plate manually. Set the wavelength of the microplate reader at 450 nm to determine the OD value of every well. (Recommend to read the OD value at the dual-wavelength 450/630 nm within 5 min).
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 Ractopamine in the sample.
7.1 Qualitative determination
The concentration range (ng/mL) can be obtained form comparing 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 0.05ppb, 1.415 for 0.15ppb, 0.74 for 0.45ppb, 0.313 for 1.35ppb, 0.155 for 4.05ppb, accordingly the concentration range of the sampleⅠ is 1.35 to 4.05ppb, and that of the sampleⅡ is 0.15ppb to 0.45ppb.
7.2 Quantitative determination
The mean values of the absorbance values obtained for 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 OD value of the sample or the standard solution
B0—the average OD value of the 0 ng/mL standard solution
Draw the standard curve with the absorption percentages of the standard solutions and the semilogarithm values of the Ractopamine 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 actual concentration of Ractopamine 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
9. Storage and expiry date
Storage: store at 2-8 ℃, not frozen.
Expiry date: 12 months; 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.