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技术文章
植物丙二醛（MDA）英文说明书-代测/价格-极威生物
点击次数：17457 发布时间：2021/1/14 22:23:55
植物丙二醛（MDA）英文说明书-代测/价格-GIVEI生物
Plant MDA
FOR RESEARCH USE ONLY
Assay range：0.2 nmol/L – 4.8 nmol/L
96 determinations
Purpose
This kit allows for the determination of MDA concentrations in Plant tissue, cell ａnd other samples.
Principle of the assay
The kit assay Plant MDA level in the sample，use Purified Plant MDA antibody to coat microtiter plate wells, make solid-phase antibody, then add MDA to wells, Combined MDA antibody which With HRP labeled,become antibody - antigen - enzyme-antibody complex, after washing Completely, Add TMB substrate solution,TMB substrate becomes blue color At HRP enzyme-catalyzed, reaction is terminated by the addition of a sulphuric acid solution ａnd the color change is measured spectrophotometrically at a wavelength of 450 nm. The concentration of Plant MDA in the samples is thendetermined by comparing the O.D. of the samples to the standard curve.
植物丙二醛（MDA）英文说明书-代测/价格-GIVEI生物
Materials provided with the kit
Materials provided with the kit
1 wash solution 20ml×1bottle 7 S Solution 6ml×1 bottle
2 HRP-Conjugate reagent 6ml×1 bottle 8 Standard（9.6nmol/L0.5ml×1 bottle
3 Microelisa stripplate 12well×8strips 9 Standard diluent 1.5ml×1bottle
4 Sample diluent 6ml×1 bottle 10 Instruction 1
5 Chromogen Solution A 6ml×1 bottle 11 Closure plate membrane 2
6 Chromogen Solution B 6ml×1 bottle 12 Sealed bags 1
Specimen requirements1. extract as soon as possible after Specimen collection,and according to therelevant literature, ａnd should be experiment as soon as possible after the
extraction. If it can’t, specimen can be kept in -20 ℃ to preserve, Avoidrepeated freeze-thaw cycles.
2. Can’t detect the sample which contain NaN3, because NaN3 inhibits HRPactive.
植物丙二醛（MDA）英文说明书-代测/价格-GIVEI生物
Assay procedure
Assay procedure
1. Dilute ａnd add sample:Dilute Original density Standard as follow table:
4.8 nmol/L 5 Standard 150µl Original density Standard+150µl Standard diluent
4.8 nmol/L 5 Standard 150µl Original density Standard+150µl Standard diluent
2.4 nmol/L 4 Standard 150µl 5 Standard+150µl Standard diluent
1.2 nmol/L 3 Standard 150µl 4 Standard+150µl Standard diluent
0.6 nmol/L 2 Standard 150µl 3 Standard +150µl Standard diluent
0.3 nmol/L 1 Standard 150µl 2 Standard +150µl Standard diluent
2.add sample：Set blank wells separately (blank comparison wells don’t addsample ａnd HRP-Conjugate reagent, other each step operation is same).testing sample well. add Sample dilution 40µl to testing sample well, then addtesting sample 10µl (sample final dilution is 5-fold), add sample to wells ,don’t touch the well wall as far as possible, ａnd Gently mix.
3.Incubate: After closing plate with Closure plate membrane ,incubate for 30
min at 37℃.
4.Configurate liquid: 30-fold(or 20-fold) wash solution diluted 30-fold (or 20-fold)with distilled water ａnd reserve.
4.Configurate liquid: 30-fold(or 20-fold) wash solution diluted 30-fold (or 20-fold)with distilled water ａnd reserve.
5.washing：Uncover Closure plate membrane, discard Liquid, dry by swing,add washing buffer to every well, still for 30s then drain, repeat 5 times, dry bypat.
6.add enzyme：Add HRP-Conjugate reagent 50µl to each well, except blankwell.
7.incubate：Operation with 3.
8.washing：Operation with 5.
9.color：Add Chromogen Solution A 50ul ａnd Chromogen Solution B to eachwell, evade the light preservation for 10 min at 37℃
10.S the reaction：Add S Solution50µl to each well, S the reaction(theblue color change to yellow color).
11.assay：take blank well as zero , Read absorbance at 450nm after AddingS Solution ａnd within 15min.
植物丙二醛（MDA）英文说明书-代测/价格-GIVEI生物
Calculate
Calculate
Take the standard density as the horizontal, the OD value for thevertical ,draw the standard curve on graph paper, Find out the correspondingdensity according to the sample OD value by the Sample curve, multiplied bythe dilution multiple, or calculate the straight line regression equation of thestandard curve with the standard density ａnd the OD value ,with the sampleOD value in the equation, calculate the sample density, multiplied by thedilution factor, the result is the sample actual density.
植物丙二醛（MDA）英文说明书-代测/价格-GIVEI生物
Important notes
Important notes
1. The kit takes out from the refrigeration environment should be balanced15-30 minutes in the room temperature, ELISA plates coated if has not useup after opened, the plate should be stored in Sealed bag.
2. washing buffer will Crystallization separation, it can be heated the waterhelps dissolve when dilute . Washing does not affect the result.
3. add Sample with sampler Each step, And proofread its accuracy frequently,avoids the experimental error. add sample within 5 min, if the number ofsample is much , recommend to use Volley .
4. if the testing material content is excessively higher (The sample OD isbigger than the first standard well ),please dilute Sample (n-fold), Pleasediluente ａnd multiplied by the dilution factor.（×n×5）.
5. Closure plate membrane only limits the disposable use, to avoidcross-contamination.
6. The substrate evade the light preservation.
7. Please according to use instruction strictly, The test result determinationmust take the microtiter plate reader as a standard.
8. All samples, washing buffer ａnd each kind of reject should according toinfective material process.
9. Do not mix reagents with those from other lots.
Storage ａnd validity
1．Storage： 2-8℃.
2．validity： six months
原创作者：上海极威生物科技有限公司
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