考试百科Human TNF-α
FOR RESEARCH USE ONLY
Assay range：：：：20 ng/L -400 ng/L 96japan life
determinations
Purpo
This kit allows for the determination of TNF-α concentrations in Human rum, cell culture supernates and other biological fluids
Principle of the assay
The kit assay Human TNF-α level in the sample, u Purified Human TNF-α antibody to coat microtiter plate wells, make solid-pha antibody, then add TNF-α to wells, Combined TNF-α 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 spectrophoto
metrically at a wavelength of 450 nm. The concentration of Human TNF-α in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Materials provided with the kit
1 wash solution 20ml×1bottle 7 Stop Solution 6ml×1 bottle
2 HRP-Conjugate reagent 6ml×1 bottle 8 Standard（800ng/L）0.5ml×1 bottle
3 Microelisa stripplate 12well×8strips 9 Standard diluent 1.5ml×1bottle
4 Sample diluent 6ml×1 bottle 10 Instruction 1石家庄教育网
工程项目质量管理五四演讲稿5 Chromogen Solution A 6ml×1 bottle 11 Closure plate membrane
2
6 Chromogen Solution B 6ml×1 bottle 12 Sealed bags 1
沙鼠
Specimen requirements
1. extract as soon as possible after Specimen collection,and according to the relevant
literature, and should be experiment as soon as possible after the extraction. If it can’t,
specimen can be kept in -20 ℃to prerve, Avoid repeated freeze-thaw cycles.
2. Can’t detect the sample which contain NaN3, becau NaN3 inhibits HRP active.
Assay procedure
1. Dilute and add sample:Dilute Original density Standard as follow table:
400 ng/L 5 Standard 150µl Original density Standard+150µl Standard diluent
200 ng/L 4 Standard 150µl 5 Standard+150µl Standard diluent
100 ng/L 3 Standard 150µl 4 Standard+150µl Standard diluent
simple cd
50 ng/L 2 Standard 150µl 3 Standard +150µl Standard diluent
25 ng/L 1 Standard 150µl 2 Standard +150µl Standard diluent
2.add sample: Set blank wells parately (blank comparison wells don’t add sample and
HRP-Conjugate reagent, other each step operation is same). testing sample well. add Sample dilution 40µl to testing sample well, then add testing 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 rerve.
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 by pat.
水壶英文
6. Add enzyme: Add HRP-Conjugate reagent 50µl to each well, except blank well.
7. Incubate: Operation with 3.音乐会英文
8. Washing: Operation with 5.
9.Color: Add Chromogen Solution A 50ul and Chromogen Solution B to each well, evade the light prervation for 15 min at 37℃
10.Stop the reaction：Add Stop Solution50µl to each well, Stop the reaction(the blue color
change to yellow color).
化学翻译11. Assay:take blank well as zero , Read absorbance at 450nm after Adding Stop Solution and
within 15min.
