|
Product Details:
Payment & Shipping Terms:
|
Sensitivity: | 0.4ppb | Incubation Temperature: | 25℃ |
---|---|---|---|
Incubation Time: | 30min—15min | Recovery Rate: | 90±15% |
Shelf Life: | 12 Months When Properly Stored | Sample Performance: | Tissue, Honey, Milk, Feed |
MOQ: | 1 Kit | ||
High Light: | Oxytetracyline elisa kit,antibiotic residue test kit,food detective test kit |
Oxytetracyline ELISA Test Kit
Catalog No. LSY-10044
1. Principle
This test kit is based on the competitive enzyme immunoassay for the detection of Oxytetracyline in the sample. The coupling antigens are pre-coated on the micro-well stripes. The Oxytetracyline in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti- Oxytetracyline 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 Oxytetracyline in it. This value is compared to the standard curve and the Oxytetracyline concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 0.4 ppb
Incubation Temperature: 25℃
Incubation Time: 30min—15min
Detection limit:
Tissue, Honey about 20ppb
Milk (method 1) about 20ppb
Milk (method 1) about 40ppb
Feed about 600ppb
Note: ppb=ng/mLor ng/g
Cross-reaction rate:
Oxytetracyline 100%
Tetracycline 200%
Chlortetracycline 240%
Doxycycline 20%
Recovery rate: 90±15%
3. Components
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:
5.1 Tissue
Fold of dilution of the sample: 20
5.2 Honey
1) Take 1± 0.01g honey sample into 10 mL centrifuge tube; Add 2ml deionized water, shake with oscillator fully for 1 min to dissolve; take 100ul dissolved solution, add 400ul diluted redissolving solution, oscillator for 10S to mix it evenly;
2) Take 50ul for analysis immediately.
Fold of dilution of the sample: 10
5.3 Milk (method 1)
1) Thaw the collected liquid milk sample, then put at room temperature for 30min;
2) Put tips in the down-layer of milk, take 1ml sample into 2ml centrifuge tube(note: do not take the up-layer cream);
3) Add 50ul 1M HCl, shake strongly for 1min(or oscillator for 30s);
4) Centrifuge at above 4000 r/min at room temperature (20 - 25 ℃) for 10 minutes;
5) Take up-layer clear liquid 50ul into another clean centrifuge tube(note: do not take the up-layer cream), add 450ul diluted redissolving solution, shake strongly for 1min(or oscillator for 30s);
6) Take 50ul for analysis immediately.
Fold of dilution of the sample: 10
Milk (method 2)
1) Take 50ul liquid sample into 1950ul diluted redissolving solution; oscillator fully for 1min evenly;
2) Take 50ul for analysis immediately.
Fold of dilution of the sample: 40
5.4 Feed (Cattle feed, pig feed)
1. Take 1± 0.01 g feed sample into 50 mL centrifuge tube, add 5 mL deionized water, shake with oscillator for 2min until feed separate completely, centrifuge at above 4000 r/min for 10 minutes.
2. Take 40ul up-layer clear liquid into a new centrifuge tube, add 1560ul diluted redissolving solution, shake with oscillator for 30s;
3. Take 50 µL for analysis.
Fold of dilution of the sample: 200
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 Oxytetracyline 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 0ppb, 1.816 for 0.4ppb, 1.415 for 1.2ppb, 0.74 for 3.6ppb, 0.313 for 10.8ppb, accordingly the concentration range of the sampleⅠis 3.6 to 10.88ppb, and that of the sample Ⅱ is 0.4 to 1.2 ppb.
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 Oxytetracyline 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 Oxytetracyline concentration in the sample.
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.