Nitrofuran(AMOZ) ELISA test kit 1.Sensitivity:0.1ppb 2.Spec.:96 wells/kit 3.Brand:Green Spring 4.ISO9001:2008
Nitrofuran(AMOZ) ELISA test kit
1.Introduction:Nitrofuran ELISA test kit is based on the competitive enzyme immunoassay for the detection of Aminohydantion (AMOZ) in the sample. The coupling antigens are pre-coated on the micro-well stripes. The Aminohydantion (AMOZ) in the sample and the coupling antigens pre-coated on the micro well stripes compete for the anti AMOZ antibodies.Aftertheadditionoftheenzymeconjugate,the TMB substrate is added for coloration. The optical density (OD) value of the sample has an egative correlation with the AHD in it. This value is compared to the standard curve and the AMOZ concentration is subsequently obtained.
2. ELISA procedures procedures1 Bring test kit to the room temperature (20 - 25 °C ) for at least 30 min, note that each reagent must be shaken evenly before use; put the required micro-well strips into plate frames. Re-sealed the unused microplate, stored at 2 - 8 °C , not frozen.2 Solution preparation: dilute 40 mL of the concentrated washing buffer (20 × concentrated) with the distilled or deionized water to 800 mL (or just to the required volume) for use.3 Numbering: number the micro-wells according to samples and standard solution, each testing sample and standard preparation should be performed in duplicate, record their positions.4 Add 50 µ L of the sample or standard solution to separate duplicate wells, an d add 50 µ L of the antibody working solution into each well, seal the microplate with the cover membrane, Mix gently by shaking the plate manually, incubate at 25 °C for 1 h.5 Pour the liquid out of micowell, flap to dry on absorbent paper, add 250 µ L/well of washing buffer to wash microplate for 10 s, repeat 4 - 5 times, then take out and flap to dry with absorbent paper .6 Add 100 µ L of the enzyme conjugate into each well; and incubate at 25 °C for 30 min.Take out microplate, continue as described in 5.7 Coloration: add 50 µ L of the substrate A solution and then 50 µ L of the B solution into each well.Mix gently by shaking the plate manually, and incubate at 25 °C for 30 min at dark for coloration.
8 Determination: add 50 µ L of the stop solution into each well. Mix gently by shaking the platemanually. Set the wavelength of the microplate reader at 450 nm to determine the OD value ofevery well.(Recommend to read the OD value at the dual-wavelength 450/630 nm within 5 min) .
3. Result judgmentThere are two methods to judge the results: the first one is the rough judgment, while the second is the quantitative determination. Note that the OD value of the sample has a negative correlation with the AOZ in the sample.3.1 Qualitative Qualitative Qualitative Qualitative determination determination determination determination.The concentration range (ng/mL) can be obtained from comparing the average OD value of the sample with that of the standard solution. Assuming that the OD value of the sample1 is 0.268,and that of the sample 2 is 1.230, the OD value of standard solution is: 1.671 for 0ppb, 1.425 for 0.05ppb, 1.103 for 0.15ppb, 0.567 for 0.45 ppb, 0.205 for 1.35 ppb ,0.104 for 4.05 ppb, accordingly the concentration range of the sample1 is 0.45 to 1.35, and that of the sample2 is 0.05 to 0.15ppb.
1 Bring all reagents and micro-well strips to balance at the room temperature (20-25 °C ) beforuse.2 Return all reagents to 2 - 8 °C immediately after use.3 The reproducibility of the ELISA analysis, to a large degree, depends on the consistency plate washing. The correct operation of plate washing is the key point in the procedures ELISA.
4 For the incubation at constant temperatures, all the samples and reagents must avoid light exposure, and each microplate should be sealed by the cover membrane.5 The room temperature below 20 °C or the temperature of the reagents and the samples being not returned to the room temperature (20 - 25 °C ) will lead to a lower standard OD value.6 Dryness of the microplate in the washing process will be accompanied by the situations including the non-linear standard curves and the undesirable reproducibility; So continue to next step immediately after washing.7 Mix evenly, otherwise there will be the undesirable reproducibility.8 The stop solution is the 2M sulfuric acid solution, avoid contacting with the skin.9 Do not use the kit exceeding its expiry date. The use of diluted or adulterated reagents from the kits will lead to the changes in the sensitivity and the detecting OD values. Do not exchange the reagents from the kits of different lot numbers to use.10 Put the unused microplate into an auto-sealing bag to re-seal it. The standard substance and the colourless color former is light sensitive, and thus they cannot be directly exposed to the light.11 Discard the colouration solution with any color that indicates the degeneration of this solution.The detecting value of the standard solution 1 ( 0 ppb ) of less than 0.5 indicates its degeneration.
Nitrofuran AMOZ ELISA Test kit 0.1 ppb