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植物(Plant)阿魏酸(FA) ELISA 检测试剂盒说明书

上海隆诚生物科技有限公司发布时间:2020/11/12 12:30:38

 
Plant Ferulic Acid (FA) ELISA Kit instruction
Intended use
This FA ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or
therapeutic procedures. The Stop Solution changes the color from blue to yellow and the intensity of the
color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of FA in
the sample, this FA ELISA Kit includes a set of calibration standards. The calibration standards are
assayed at the same time as the samples and allow the operator to produce a standard curve of Optical
Density versus FA concentration. The activities of FA in the samples is then determined by comparing the
O.D. of the samples to the standard curve.
Sample collection and storages
1. Can’t detect the samples which contain NaN3, because NaN3 inhibits HRP activity of the horseradish
peroxidase.
2. Extract as soon as possible after Specimen collection, Extracted according to the relevant literature.
Cell culture supernates and plant exact fluids - Remove particulates by centrifugation and assay
immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw.
Materials required but not supplied
1. Standard microplate reader(450nm)
2. Precision pipettes and Disposable pipette tips.
3. 37 incubator
Precautions
1.
Do not substitute reagents from one kit to another. Standard, conjugate and microplates are
matched for optimal performance. Use only the reagents supplied by manufacturer.
2. Do not remove microplate from the storage bag until needed. Unused strips should be stored at
2-8°C in their pouch with the desiccant provided.
3. Mix all reagents before using.
Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)热线:021-51172858
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Materials supplied
Name
96 determinations
48 determinations
Microelisa stripplate
12*8strips
12*4strips
Standard
0.3ml*6tubes
0.3ml*6tubes
Sample Diluent
6.0ml
3.0ml
HRP-Conjugate reagent
10.0ml
5.0ml
20X Wash solution
25ml
15ml
Chromogen Solution A
6.0ml
3.0ml
Chromogen Solution B
6.0ml
3.0ml
Stop Solution
6.0ml
3.0ml
Closure plate
membrane
2
2
User manual
1
1
Sealed bags
1
1
Note: Standard (S0 → S5) concentration was followed by: 0,0.5,1,2,4,8 ppb
Reagent preparation
20×wash solution:Dilute with Distilled or deionized water 1:20.
Assay procedure
1. Prepare all re a g e n ts before starting assay procedure. It is recommended that all Standards and
Samples be added in duplicate to the Microelisa Stripplate.
2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.
3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well; Blank
well doesn’t add anyting.
4. Add 100μl of HRP-conjugate reagent to each well, cover with an ccresive strip and incubate for 60
minutes at 37°C.
5. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by
filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher.
Complete removal of liquid at each step is essential to good performance. After the last wash, remove
any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper热线:021-51172858
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towels.
6.
Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and
incubate for 15 minutes at 37°C. Protect from light.
7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the
color in the wells is green or the color change does not
appear uniform, gently tap the plate to ensure thorough mixing.
8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.
Calculation of results
1. This standard curve is used to determine the amount in an unknown sample. The standard curve is
generated by plotting the average O.D. (450 nm) obtained for each of the six standard
concentrations on the vertical (Y) axis versus the corresponding concentration on the horizontal (X)
axis.
2. First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by
the mean value of the zero standard before result interpretation. Construct the standard curve
using graph paper or statistical software.
3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a
horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis
and read the corresponding concentration.
4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit
age can cause variation in result. Each user should obtain their own standard curve.
5. The sensitivity by this assay is 0.1 ppb
6. Standard curve热线:021-51172858
Storage2-8.
validitysix months.
FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE
READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
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阅读人数:271 原创作者:上海隆诚生物科技有限公司

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