Tissue, honey, acquatic product
Nitrofuran (AHD) ELISA Test Kit
1. Principle
This test kit is based on the competitive enzyme immunoassay for the detection of Aminohydantion (AHD) in the sample. The coupling antigens are pre-coated on the micro-well stripes. The Aminohydantion (AHD) in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti-AHD 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 AHD in it. This value is compared to the standard curve and the AHD concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 0.1 ppb
Detection limit
Tissue and honey·········································································· 0.2 ppb
Shrimp and fish (some interference in shrimp and fish)··················· 0.3 ppb
Recovery rate
Shrimp and fish············································································ 90±15%
Honey, chicken meat /liver····························································· 80±15%
Cross-reaction rate
AHD································································································ 100%
AMOZ····························································································· ?0.1%
AOZ································································································ ?0.1%
SEM······························································································· ?0.1%
3. Components
1) Micro-well strips: 12 strips with 8 removable wells each
2) 6× standard solution (1 mL each): 0 ppb, 0.1 ppb, 0.3 ppb, 0.9 ppb, 2.7 ppb and 8.1 ppb
3) Enzyme conjugate (12 mL)····························································· red cap
4) Antibody working solution (7 mL)·················································· blue cap
5) Substrate A solution (7 mL)························································· white cap
6) Substrate B solution (7 mL)························································ black cap
7) Stop solution (7 mL)································································· yellow cap
8) 20× concentrated washing buffer (40 mL)···································· white cap
9) 2× concentrated redissolving solution (50 mL)···················· transparent cap
10) 2-Nitrobenzaldehyde (15.1 mg)···················································· white cap
4. Materials required but not provided
1) Equipments: microplate reader, printer, homogeniser, nitrogen-drying device, vortex, centrifuge, measuring pipets, and balance (a sensibility reciprocal of 0.01 g);
2) Micropipettors: single-channel 20 to 200 µL and 100 to 1000 µL, and multi-channel 250 µL;
3) Reagents: methanol, NaOH, ethyl acetate, n-Hexane, HCI (approx36.5%), K2HPO4·3H2O, 2-Nitrobenzaldehyde (C7H5NO3), K2Fe(CN)5NO·3H2O and ZnSO4·7H2O
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) 10 mM 2-Nitrobenzaldehyde solution: dissolve 15.1 mg 2-Nitrobenzaldehyde in 10 mL methanol (concentration: 10 mM).
2) the 2×concentrated redissolving solution is mixed with deionized water at 1:1 (1 mL concentrated redissolving solution + 1 mL deionized water), used for sample redissolving.
3) C solution (for milk sample): dissolve 12.5 g K2Fe(CN)5NO·3H2O in deionized water to 100 mL.
4) D solution (for milk sample): dissolve 29.8 g ZnSO4·7H2O in deionized water to 100 mL.
5) 0.1 M K2HPO4: dissolve 22.8 g K2HPO4·3H2O in deionized water to 1 L.
6) 1 M HCl: dissolve 8.6 mL HCI (approx 36.5%) in water to 100 mL.
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