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Sensitivity: | 0.2ppb | Incubation Temperature: | 25℃ |
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Incubation Time: | 30min—15min | Sample Performance: | Milk,honey, Tissue |
Recovery Rate: | 90%±30% | MOQ: | 1 Kit |
Shelf Life: | 12 Months When Properly Stored | ||
High Light: | Erythromycin ELISA kit,antibiotic residue test kit,honey test kit |
Erythromycin ELISA Test Kit
Catalog No. LSY-10050
1. Principle
This test kit is based on the competitive enzyme immunoassay for the detection of Erythromycin in the sample. The coupling antigens are pre-coated on the micro-well stripes. The Erythromycin in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti- Erythromycin 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 Erythromycin in it. This value is compared to the standard curve and the Erythromycin concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 0.2 ppb
Incubation Temperature: 25℃
Incubation Time: 30min—15min
Detection limit:
Raw milk about 10ppb
Honey (method 1) about 4ppb
Honey (method 2) about 0.2ppb
Tissue about 15ppb
Note: ppb=ng/mL or ng/g
Cross-reaction rate:
Erythromycin 100%
Recovery rate:
90%±30%
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 Milk
Fold of dilution of the sample: 20
5.2 Honey (Method 1)
1) Take 1.0±0.05g honey sample into 50ml Polystyrene centrifuge tube;
2) Add 2ml Methanol-2% NaCl Solution, vortex for 2min until honey is dissolved completely, Centrifuge at 3000g at room temperature (20 - 25 ℃) for 5min;
3) Take 100ul bright solution into 2ml Polystyrene centrifuge tube, add 900ul redissolving solution;
4) Take 50 µL for analysis.
Fold of dilution of the sample: 20
5.2 Honey (Method 2)
1) Take 2.0±0.05g honey sample into 50ml Polystyrene centrifuge tube;
2) Add 2ml 0.1M CB solution, vortex until honey sample is completely dissolved, then add 6ml Ethyl acetate, shake for 5min;
3) Centrifuge at 3000g at room temperature (20 - 25 ℃) for 5min;
4) Take 3ml up-layer organic phase into 10ml clean dry glass tube, blow to dry in 50-60℃ water bath by Nitrogen;
5) Add 0.5ml redissolving solution, vortex for 5min, mix it evenly;
6) Take 50 µL for analysis.
Fold of dilution of the sample: 0.5
5.3 Tissue
1) Take 1.0±0.05g tissue sample into 50ml Polystyrene centrifuge tube;
2) Add 4ml 0.05M NaOH solution, vortex for 5min until tissue sample separate;
3) Centrifuge at 4000g at room temperature (20 - 25 ℃) for 5min;
4) Take 200ul up-layer clear liquid, add 750ul redissolving solution and 50ul 0.1M HCl Solution, vortex for 1min and mix evenly(If the sample is too cloudy, please centrifuge and analyze);
5) Take 50 µL for analysis.
Fold of dilution of the sample: 25
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
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 Erythromycin 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 Erythromycin 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.