Penicillin ELISA 试剂盒
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北京 101111
Quick Overview for Penicillin ELISA 试剂盒 (ABIN997088)
抗原
See all Penicillin products适用
检测方法
实验类型
应用范围
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原理
- Enzyme Immunoassay for the Quantitative Determination of Penicillin in Food
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Analytical Method
- Quantitative
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灵敏度
- 3 ng/mL
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实验时间
- 2 - 3 h
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板类型
- Pre-coated
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实验流程
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- Prepare samples as described above.
2. Pipet 100 μL ready-to use standards or prepared samples in duplicate into the appropriate wells of the microtiter plate. Immediately add 50 μL penicillin antibody into each well.
3. Cover the microtiter plate with a plastic foil and incubate for 60 minutes at room temperature on a microtiter plate shaker (or 90 minutes without shaker).
4. Wash the plate three times as follows: Discard the contents of the wells (dump or aspirate). Pipet 300 μL of diluted washing solution into each well. After the third repetition empty the wells again and remove residual liquid by striking the plate against a paper towel. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbencies.
5. Pipet 100 μL of conjugate (anti-mouse-IgG-HRP) into each well.
6. Cover the microtiter plate with a plastic foil and incubate for 60 minutes at room temperature on a microtiter plate shaker (or 90 minutes without shaker).
7. Wash the plate as outlined in 4.
8. Pipet 100 μL of substrate solution into each well.
9. Allow the reaction to develop in the dark (e.g. cupboard or drawer, the chromogen is light-sensitive) for 20 minutes at room temperature.
10. Stop enzyme reaction by adding 100 μL of stop solution (0.5 M H2SO4) into each well. The blue colour will turn yellow upon addition.
11. After thorough mixing, measure absorbance at 450 nm (reference wavelength 620 nm), using an ELISA reader. The colour is stable for 30 minutes.
- Prepare samples as described above.
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结果分析
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- Calculate the average optical density (OD 450 nm) for each set of reference standards or samples.
2. Construct a standard curve by plotting the mean optical density obtained for each reference standard against its concentration in ng/mL on semi-log graph paper with the optical density on the vertical (y) axis and the concentration on the horizontal (x) axis.
3. Using the mean optical density value for each sample, determine the corresponding concentration ' of penicillin in ng/mL from the standard curve. Depending on experience and/or the availability of ' computer capability, other methods of data reduction may be employed.
4. The diluted samples must be further converted by the appropriate dilution factor. The dilution factor is 20 for shrimps und 4 for milk extraction according to the sample preparation procedure as described above.
TYPICAL STANDARD VALUES
The following table contains an example for a typical standard curve. The binding is calculated as percent of the absorption of the 0 ng/mL standard. These values are only an example and should not be used instead of the standard curve which has to be measured in every new test. Penicillin (ng/mL) ( % binding of 0 ng/mL) 0 100 4 85 10 70 40 35 100 15 400 5
- Calculate the average optical density (OD 450 nm) for each set of reference standards or samples.
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限制
- 仅限研究用
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储存条件
- 4 °C
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储存方法
- Store at 2-8 °C
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- Penicillin
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物质类
- Chemical
抗原 See all Penicillin products
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