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Sulfamethazine (SM2) ELISA Kit

Reactivity: Chemical Colorimetric Competition ELISA
Catalog No. ABIN400606
  • Target
    Sulfamethazine (SM2)
    Reactivity
    Chemical
    Detection Method
    Colorimetric
    Method Type
    Competition ELISA
    Application
    ELISA
    Purpose
    The test kit is based on the competitive enzyme immunoassay for the detection of Sulfamethazine(SM2) in the chicken, pork, milk, honey and egg. The coupling antigens are pre-coated on the micro-well stripes. The Sulfamethazine in the sample and the conjugated antigen pre-coated on the micro-well stripes compete for the anti-Sulfamethazine 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 Sulfamethazine concentration in the sample. This value is compared to the standard curve and the Sulfamethazine concentration is subsequently obtained.
    Analytical Method
    Qualitative and Quantitative
    Components
    Micro-well strips: 12 strips with 8 removable wells each 6 standard solution (1 mL each): 0 ppb, 1 ppb, 3 ppb, 9 ppb, 27 ppb and 81 ppb, Enzyme conjugate (7 mL) red cap, Antibody working solution (10 mL) blue cap, Substrate A solution (7 mL) white cap, Substrate B solution (7 mL) black cap, Stop solution (7 mL) yellow cap, 20 concentrated washing buffer (40 mL) white cap, 2 concentrated redissolving solution (50 mL) transparent cap
    Material not included
    Equipments: microplate reader, printer, mixer or stomacher, nitrogen-drying device, oscillator, centrifuge, measuring pipets, balance( a reciprocal sensibility of 0.01 g), Micropipettors: single-channel 20-200 L and 100-1000 L, and multi-channel 250 L, 3) Reagents: Acetonitrile(CH3CN), ethyl acetate, N-hexane, Na2HPO412H2O, NaH2PO42H2O, NaCl
  • Plate
    Pre-coated
    Protocol
    Sample pre-treatment: Instructions (The following points must be dealt with before the pre-treatment) 1)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: 1 2 M NaCI: dissolve 11.69 g NaCI in the deionized water to 100 mL. 2 0.02 M PB buffer: weight 2.58 g Na2HPO412H2O and 0.44 g NaH2PO42H2O, dissolve in the deionized water to 500 mL. 3 Acetonitrile(CH3CN)-water solution: ?CH3CN:?H2O =84:16. 4 The 2 concentrated redissolving solution is mixed with deionized water at 1:1 (1 mL concentrated redissolving solution + 1 mL deionized water), for the sample redissolving. 5.1 High-detection-limit samples 5.1.1 Animal tissues (meat, liver), shrimp, fish, egg 1 Take the sample, homogenize at 10000 r/min for 1 min. 2 Weigh 30.05 g of the homogenized sample, put into centrifuge tube, add 9 mL of the CH3CN-water solution, shake properly for 10 min, centrifuge at above 4000 r /min at 15 ? for 10 min. 3 Transfer 4 mL of the supernatant into a new vessel, add 2 mL of 2 M NaCI solution and 7 mL ethyl acetate, shake for 10 min, and centrifuge at above 4000 r/min at 15 ? for 5 min. Transfer the supernatant into a new vessel, blow to dry with nitrogen at 50oC. Add 1 mL of the diluted redissolving solution, shake for 1 min, add 1 mL N-hexane, mix for 2 min and centrifuge at 4000 r/min at 15 ? for 5 min, remove the upper layer. Take 20 L of the lower for further analysis. Fold of dilution of the sample: 1 It needs five fold dilution of the sample(1 mL sample+4 mL of the diluted redissolving solution) if the detection is based on the most residue (100 ppb) of national regulation. 5.2 Low-detection-limit samples 5.2.1 Animal tissues (meat, liver) 1) Weight 2.0 0.05 g of the sample, add 10 mL 0.02 M PB buffer, shake upside down for 10 min, put into 37oC constant temperature container for 30 min, centrifuge at above 5000 r/min at 10oC for 10 min. 2) Take 20 L of the clear supernatant (upper layer) for further analysis. Fold of dilution of the sample: 5 Detection limit: 5 ppb 5.2.2 Animal tissues (meat or chicken) 1 Weigh 2.0 0.05 g of the sample, add 10 mL of 0.02 M PB buffer and 5 mL n-hexane, shake upside and down for 10 min, centrifuge at above 5000 r/min at 10oC for 10 min. 2 Remove n-hexane phase (upper layer), take 100 L of the lower, add 100 L 0.02 M PB buffer, mix properly. 3 Take 20 L for analysis. Fold of dilution of the sample: 10 Detection limit: 10 ppb 5.2.3 Serum 1 Place sample at room temperature for 30 min, centrifuge at above 4000 r/min at 10oC for10 min, separate or filter serum. 2 Take 1 mL serum, add 3 mL 0.02 M PB buffer, mix properly. 3 Take 20 L for further analysis. Fold of dilution of the sample: 4 Detection limit: 4 ppb 5.2.4 Honey 1 Put 1.0 0.05 g honey into 50 mL centrifuge tube, add 2 mL 0.02 M PB buffer to dissolve. 2 Add 8 mL of the ethyl acetate, shake for 10 min, centrifuge at above 4000 r/min at room temperature (20-25?) for 10 min. 3 Take the upper layer, blow to dry with nitrogen at 50oC, add 0.5 mL of diluted redissolving solution to redissolve. 4 Take 20 L for further analysis. Fold of dilution of the sample: 1 5.2.5 Urine 1 Add 3 mL 0.02 M PB buffer and 1 mL of the centrifuged clear sample, mix properly. 2 Take 20 L for further analysis. Fold of dilution of the sample: 4 Detection limit: 4 ppb Milk Take 1 mL milk, add 0.02 M PB buffer, dilute at 1:20(?/?) (20 L milk + 380 L of 0.02 M PB buffer). Take 20 L for further analysis. Fold of dilution of the sample:20
    Restrictions
    For Research Use only
  • Storage
    4 °C
  • Target
    Sulfamethazine (SM2)
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