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Product Details:
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Sensitivity: | 0.1ppb | Incubation Temperature: | 25℃ |
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Incubation Time: | 30min—15min | Sampleperformance: | Honey |
Shelf Life: | 12 Months When Properly Stored | Specifications: | 96 Wells/kit |
MOQ: | 1 Kit | ||
High Light: | antibiotics residue test kit,TCS elisa kit,antibiotic residue elisa diagnostic kits |
Tetracyclines(TCs) ELISA Test Kit
Catalog No. LSY-10006-2
1. Principle
This test kit is based on the competitive enzyme immunoassay for the detection of Tetracyclines in the honey sample. The coupling antigens are pre-coated on the micro-well stripes. The Tetracyclines in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti-Tetracyclines antibody. 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 Tetracyclines in it. This value is compared to the standard curve and the Tetracyclines concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 0.1 ppb
Incubation Temperature: 25℃
Incubation Time: 30min—15min
Detection limit:
Honey 4ppb
Cross-reaction rate:
Doxycycline 100%
Tetracycline 150%
Minocycline 92%
Pyrithione 76%
Chlortetracycline 75%
Demethylchromycin 70%
Oxytetracyline 83%
Recovery rate:
Honey 70%~120%
3. Components
1 | Micro-well strips |
12 strips with 8 removable wells each |
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2 | 6× standard solution (1mL each) | 0ppb | 0.1ppb |
0.3ppb | 0.9ppb | ||
2.7ppb | 8.1ppb | ||
3 | Enzyme conjugate | 7ml | red cap |
4 | Antibody working solution | 7ml | blue cap |
5 | Substrate A | 7ml | white cap |
6 | Substrate B | 7ml | black cap |
7 | Stop solution | 7ml | yellow cap |
8 | 20× concentrated washing buffer | 15ml | white cap |
9 | 20× sample extract A | 15ml | yellow cap |
10 | 2× sample extract B | 50ml*2 | transparent cap |
11 | 20× sample diluent | 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 of any kind of sample:
1) Only the disposable tips can be used for the experiments and the tips must be changed when used for absorbing different reagents;
2) Before the experiment, each experimental equipment must be clean and should be re-cleaned if necessary, in order to avoid the contamination that interferes with the experimental results.
Solution preparation before sample pre-treatment:
1) Sample extract A
1 part of 20× sample extract A + 19 parts of deionized water
2) Sample extract B
1 part of 2× sample extract B + 1 part of deionized water
3)1M NaOH solution
Weigh 4g NaOH, add deionized water to 100ml
4) Sample diluent
1 part of 20× sample diluent + 19 parts of deionized water
5.1 Honey
1) Take 2± 0.05 g of the honey sample into 50 mL centrifuge tube, add 3mL diluted Sample extract A, shake for 3min;
2) Then add 600ul 1M NaOH solution and 2.4ml diluted Sample extract B, shake for 3min; centrifuge at above 4000 r/min at room temperature (20 - 25 ℃) for 5 minutes.
3) Take 50ul up-layer clear liquid, add 450ul diluted Sample diluent, mix it evenly;
4) Take 50ul above mixed solution for analysis.
Fold of dilution of the sample: 40
6. ELISA procedures
Instructions
1. Bring all reagents and micro-well strips to the room temperature (20-25 ℃).
2. Return all reagents to 2-8 ℃ immediately after use.
3 .The reproducibility of the ELISA analysis, to a large degree, depends on the consistency of plate washing. The correct operation of plate washing is the key point in the procedures of ELISA.
4. For the incubation at constant temperatures, all the samples and reagents must avoid light exposure, and each microplate should be sealed by the cover membrane.
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 Tetracyclines in the sample.
7.1 Qualitative determination
The concentration range (ng/mL) can be obtained from the comparison the average OD value of the sample with that of the standard solution. Assuming that the OD value of the sample Ⅰ is 0.3, and that of the sample Ⅱ is 1.0, while those of the standard solutions are as the followings: 2.243 for 0 ppb, 1.816 for 0.1 ppb, 1.415 for 0.3 ppb, 0.74 for 0.9 ppb, 0.313 for 2.7 ppb and 0.155 for 8.1 ppb, accordingly the concentration range of the sampleⅠis 2.7 to 8.1ppb, and that of the sample Ⅱ is 0.3 to 0.9ppb.
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 0 ng/mL standard solution
Draw the standard curve with the absorption percentages of the standard solutions and the semilogarithm values of the Tetracyclines 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, thus finally obtaining Tetracyclines 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
9. Storage and expiry date
Storage: store at 2-8 ℃, not frozen.
Expiry date: 12 months; date of production is on the 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.