LSY-10064 Ofloxacin ELISA Kit 0.1ppb

Ofloxacin ELISA Kit

Catalog No. LSY-10064

1. Principle

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

2. Technical specifications

Sensitivity: 0.1 ppb

Incubation Temperature: 25℃

Incubation Time: 30min—15min

Detection limit:

Ofloxacin (OFL) 0.3ppb

Cross-reaction rate:

Ofloxacin (OFL) 100%

Recovery rate

Tissue 90±30%

3. Components

4. Materials required but not provided

• Equipments: microplate reader, printer, homogenizer, nitrogen-drying device, vortex, centrifuge, measuring pipets, balance (a sensibility reciprocal of 0.01 g)
• Micropipettors: single-channel 20-200 µL, 100-1000 µL, and multi-channel 250 µL;
• Reagents: Acetonitrile (CH₃CN), N-hexane

5. Sample pre-treatment

Instructions (The following points must be dealt with before the pre-treatment)

1) This test kit can detect tissue sample: animal tissue, poultry, aquatic. Eg: Chicken, duck, bovine, rabbit, fish, shrimp etc.

• 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 utensil 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:

• The 2× concentrated redissolving solution is diluted with deionized water at 1:1 (1 part of concentrated redissolving solution + 1 part of deionized water), used for the sample redissolving;
• Sample extracting buffer: dilute 10x sample extracting buffer with deionized water at 1:9;
• Acetonitrile (CH₃CN)-Sample extracting buffer mixing solution: mix Acetonitrile (CH₃CN) with Sample extracting buffer at 9:1 [like 90ml of Acetonitrile (CH₃CN)+ 10ml of Sample extracting buffer]

5.1 Tissue

1) Weigh 2.0 ± 0.05 g of the homogenized tissue sample into 50 ml centrifuge tube;

2) Add 6 ml of the Acetonitrile (CH₃CN)-Sample extracting buffer mixing solution, shake for 3 min, then add 3ml N-hexane, shake for 1min, centrifuge at above 4000 r/min at 15 ℃ for 5 min;

3) Take 1.5 ml of the organic phase (middle-layer) into a dry tube, blow to dry with nitrogen completely by rotary evaporation at 56 ℃

• Dissolve the dry residues in 1 mL of the N-hexane, then add 1 mL of the diluted redissolving solution, mix for 15 seconds; centrifuge at above 4000 r/min at 15℃ for 5 min.
• Discard the upper layer, take 50µl lower layer solution for further analysis.

Fold of dilution of the sample: 2