LSY-10065 Trimethoprim ELISA Test Kit

Trimethoprim ELISA Test Kit

Catalog No. LSY-10065

1. Principle

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

2. Technical specifications

Sensitivity: 0.5 ppb

Incubation Temperature: 25℃

Incubation Time: 30min—15min

Detection limit:

Tissue 5ppb

Cross-reaction rate:

Trimethoprim 100%

Sulfamerazine ＜ 0.1%

Sulfaquinoxaline <0.1%

Sulfamethazine ＜ 0.1%

Sulfamonomethoxine <0.1%

Sulfamethoxazole <0.1%

Sulfathiazole <0.1%

Sulfamethoxypyridazine <0.1%

Sulfaguanidine <0.1%

Sulfadimethoxine <0.1%

Sulfadiazine <0.1%

Sulfameter <0.1%

Recovery rate:

Tissue 60%～120%

3. Components

4. Materials required but not provided

• Equipments: microplate reader, homogenizer, oscillator, centrifuge, balance (a sensibility reciprocal of 0.01 g), graduated pipette, incubator.
• Micropipettors: single-channel 20~200 µL and 100~1000 µL, and multi-channel 30~300 µL;
• Reagents: NaOH.

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:
1) Sample extract A

1 part of 20× sample extract A + 19 parts of deionized water

2) Sample extract B

1 part of 2× sample extract B + 1 part of deionized water

3)1M NaOH solution

Weigh 4g NaOH, add deionized water to 100ml

4) Sample diluent

1 part of 10× sample diluent + 9 parts of deionized water

5.1 Tissue

1) Take 2± 0.05 g of the homogenized tissue sample into 50 mL centrifuge tube, add 3mL diluted Sample extract A, shake for 3min;

2) Then add 600ul 1M NaOH solution and 2.4ml diluted Sample extract B, shake for 3min; centrifuge at above 4000 r/min at room temperature (20 - 25 ℃) for 5 minutes.

3) Take 200ul up-layer clear liquid, add 300ul diluted Sample diluent, mix it evenly;

4) Take 50ul above mixed solution for analysis.

Fold of dilution of the sample: 10