LSY-10060 Avermectin ELISA Test Kit for milk and tissue

Avermectin ELISA Test Kit

Catalog No. LSY-10060

1. Principle

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

2. Technical specifications

Sensitivity: 0.5ppb

Detection limit

Tissue 4ppb

Liquid milk 5ppb

Note: ppb= ng/ml or ng/g

Cross-reaction rate

Avermectin 100%

Ivermectin 25%

Eprinomectin 10%

Doramectin 6%

Recovery rate

90±30%

3. Components

• Micro-well strips: 12 strips with 8 removable wells each
• 6× standard solution (1mL each): 0ppb,0.5ppb,1.5ppb, 4.5ppb,13.5ppb,40.5ppb
• 11X Concentrated Enzyme conjugate (0.7 mL)
• Enzyme conjugate dilution (7 mL)
• Substrate A (7 mL)
• Substrate B (7 mL)
• Stop solution (7 mL)
• 20× concentrated washing buffer (30 mL)
• Redissolving solution (50 mL)
• Tissue sample treatment solution (10mL)

4. Materials required but not provided

• Equipments: microplate reader (450nm, 630nm), printer, homogenizer, nitrogen-drying device, vortex, shaker, centrifuge (4000g and above), measuring pipets, balance (a reciprocal sensibility of 0.01 g), incubator (25℃), timer;
• Micropipettors: single-channel 20-200 µL, 100-1000 µL, and eight-channel 30～300 µl;
• Reagents (AR): Methanol, 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 Tissue

1) Take 2.0±0.05g homogenized tissue sample into a 10ml Polystyrene centrifuge tube;

2) Add 2ml Methanol, 100ul Tissue sample treatment solution accordingly, vortex for 5min until tissue sample separate;

3) Centrifuge at 4000g at room temperature (25±2℃) for 10min;

4) Take 200ul up-layer clear liquid, add 400ul redissolving solution, vortex for 10S and mix evenly;

5) Take 50 µL for analysis.

Fold of dilution of the sample: 6

5.2 Milk

1) Take 1ml milk sample into a 10ml Polystyrene centrifuge tube;

2) Add 3ml Methanol, vortex for 1min;

3) Centrifuge at 4000g at room temperature (25±2℃) for 10min;

4) Take 200ul up-layer liquid, add 400ul redissolving solution, vortex for 10S and mix evenly;

• Take 50 µL for analysis.

Fold of dilution of the sample: 10