Sulfonamides(SAs) ELISA Kit

Catalog No. ABIN400611

Product Details

Target: Sulfonamides(SAs)

Reactivity: Chemical

Detection Method: Colorimetric

Method Type: Competition ELISA

Application: ELISA

Purpose: This test kit is based on the competitive enzyme immunoassay for the detection of Sulfonamides in the chicken, pork, milk, honey, egg, etc. The coupling antigens are pre-coated on the micro-well stripes. The Sulfonamides in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti-Sulfonamides 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 Sulfonamides in the sample. This value is compared to the standard curve and the Sulfonamides 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, 0.4 ppb, 1.2 ppb, 3.6 ppb, 10.8 ppb and 32.4 ppb, Enzyme conjugate (7 mL) red cap, Antibody working solution (7 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, homogenizer, nitrogen-drying device, votex, centrifuge, measuring pipets, balance (a sensibility reciprocal of 0.01 g) Micropipettors: single-channel 20-200 L, 100-1000 L, and multi-channel 250 L, Reagents: Acetonitrile (CH3CN), ethyl acetate, N-hexane, Na2HPO412H2O, NaH2PO42H2O, NaOH, HCl

Application Details

Plate: Pre-coated

Protocol: Sample pre-treatment: Instructions (The following points must be dealt with before the pre-treatment) 1) This product can detect the following tissue samples:animal tissue,poultry,and aquatic tissue, eg:chicken,duck, beef,rabbit,fish,shrimp,etc. 2) Only the disposable tips can be used for the experiments and the tips must be changed when used for absorbing different reagents, 3) 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: 0.2 M NaOH: dissolve 0.8 g NaOH in deionized water to 100 mL. 0.5 M HCl (for honey): dissolve 4.3 mL HCI (36%) in deionized water to 100 mL. 0.02 M PB buffer: dissolve 2.58 g Na2HPO412H2O and 0.44 g NaH2PO42H2O in the deionized water to 500 mL. (for low-detection-limit samples) The 2 concentrated redissolving solution is diluted with deionized water at 1:1 (1 mL concentrated redissolbing solution + 1 mL deionized water), used for the treated sample redissolving. 5.1 Tissues(meat,chicken,duck, liver, shrimp, fish, egg. etc) Homogenize the sample at 10000 r/min for 1 min Weigh 3 0.05 g of the homogenized sample, put into centrifugal tube, add 3 mL 0.02 M PB buffer, shake properly. Then add 4 mL ethyl acetate and 2 mL Acetonitrile (CH3CN), shake properly for 5 min, centrifuge at above 4000 r/min at 15 ? for 10 min, Transfer 2 mL supernatant(approx 1 g sample) into a new centrifugal tube, blow to dry with nitrogen or air completely by rotary evaporation at 56oC Add 1 mL N-hexane, then add 1 mL of the diluted redissolving solution, shake strongly for 30s. Centrifuge at 4000 r/min at room temperature for 5 min, remove the upper layer Take 20 L of lower for further analysis. Fold of dilution of the sample: 1 It needs five fold to dilute the sample(1mL sample+4 mL of the diluted redissolving solution) if the detection is stipulated in the most residue (100 ppb) of national regulation. 5.2 Serum Place blood sample at room temperature (20-25oC) for 30 min, centrifuge at above 4000 r/min at 10oC for 10 min, separate or filter serum Take 1 mL serum, add 3 mL 0.02 M PB buffer, mix properly for 30s Take 20 L for further analysis Fold of dilution of the sample: 4 Detection limit: 1.6 ppb 5.3 Honey Weight 1.0 0.05 g honey into 50 mL centrifugal tube, then add 1 mL 0.5 M HCI. Be static at 37oC for 30 min. Add 2.5 mL 0.2 M NaOH (adjust pH to 5), then add 4 mL ethyl acetate, shake for 5 min, centrifuge at above 4000 r/min at room temperature (20-25oC) for 10 min. Take 2 mL supernatant, blow to dry with nitrogen at 56oC, add 0.5 mL of the diluted redissolving solution, redissolve it for 30s Take 20 L for further analysis Fold of dilution of the sample: 1 Detection limit: 1 ppb 5.4 Urine Add 3 mL 0.02 M PB buffer and 1 mL of the centrifuged clear urine sample, mix properly for 30s Take 20 L for further analysis Fold of dilution of the sample: 4 Detection limit: 1.6 ppb 5.5 Milk Take 1 mL milk, add 0.02 M PB buffer, dilute at 1:19(?/?) (20 L milk + 380 L 0.02 M PB buffer) Take 20 L for further analysis Fold of dilution of the sample: 20

Restrictions: For Research Use only

Handling

Storage: 4 °C

Target Details

Target: Sulfonamides(SAs)

Target Type: Chemical