LSY-10061 Dimitridazole ELISA Test Kit for aquatic products

Dimitridazole ELISA Test Kit

Catalog No.: LSY-10061

1. Principle

This test kit is based on the competitive enzyme immunoassay for the detection of Dimitridazole in the sample. The coupling antigens are pre-coated on the micro-well stripes. The Dimitridazole in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti- Dimitridazole 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 Dimitridazole in it. This value is compared to the standard curve and the Dimitridazole concentration is subsequently obtained.

2. Technical specifications

Sensitivity: 0.05ppb

Detection limit

Pork, Chicken, Fish, Shrimp about 0.25ppb

Note: ppb= ng/ml or ng/g

Cross-reaction rate

Dimetridazole 100%

Metronidazole 50%

Hydroxyl Nitroimidazoles 25%

Hydroxymethyl dimetridazole 25%

Tinidazole 1%

Ornidazole 2.5%

Recovery rate

90±30%

3. Components

• Micro-well strips: 12 strips with 8 removable wells each
• 6× standard solution (1mL each): 0ppb, 0.05ppb, 0.15ppb, 0.45ppb, 1.35ppb and 4.05ppb
• Enzyme conjugate (12 mL) 1 bottle
• Antibody working solution (7mL) 1 bottle
• Substrate A solution (7 mL) 1 bottle
• Substrate B solution (7 mL) 1 bottle
• Stop solution (7 mL) 1 bottle
• 20× concentrated washing buffer (30 mL) 1 bottle
• Redissolving solution (50 mL) 1 bottle

4. Materials required but not provided

• Equipments: microplate reader (450nm, 630nm), printer, homogenizer, nitrogen-drying device, vortex, shaker, centrifuge (3000g and above), measuring pipets, balance (a reciprocal sensibility of 0.01 g), incubator (4℃,25℃), water bath, timer;
• Micropipettors: single-channel 20-200 µL, 100-1000 µL, and eight-channel 30～300 µl;
• Reagents (AR): Ethyl acetate, n-Hexane, Deionized water.

5. Sample pre-treatment

Instructions

The following points must be dealt with before the pre-treatment of any kind of sample:

• Only the disposable tips can be used for the experiments and the tips must be changed when used for absorbing different reagents;
• 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:

• Washing buffer: 1 part 20× concentrated washing buffer + 19 parts deionized water.

5.1 Samples preparation

a) Tissue (Chicken, pork, fish, shrimp, seafood etc.)

1) Weigh 2 g of the homogenized sample (tissue) into 50ml plastic centrifuge tube;

2) Add 2ml deionized water, shake strongly for 1min (or use vortex for 30s);

3) Add 8ml Ethyl acetate, shake or vortex for 3min (Note: if there is Shaker in lab, shake for 1min, then put it in Shaker at 300rpm at 25℃ for 10min);

4) Centrifuge at above 3000 g at room temperature for 5min;

5) Take 2ml supernatant into a clean glass centrifuge tube;

6) Blow to dry in 50-60℃ water bath by nitrogen-drying device;

7) Add 0.5ml n-Hexane, then 0.5ml Redissolving solution, shake up and down for 20 times;

8) Centrifuge at above 3000 g at room temperature for 5min;

9) Discard up-layer n-Hexane and middle-layer Impurity layer;

10) Take 50ul down-layer liquid to test.

Fold of dilution of the sample: 1