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更新時間:2014-10-11 14:47:08瀏覽次數(shù):144次
聯(lián)系我時,請告知來自 環(huán)保在線豬褪黑素(MT)ELISA 試劑盒
規(guī) 格:96T/48T
檢測標本:血清,血漿,尿液,胸腹水,腦脊液,細胞培養(yǎng)上清,組織勻漿等
檢測方法:ELISA
檢測類型:酶聯(lián)免疫夾心法
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Drug Names豬褪黑素(MT)ELISA 試劑盒
Generic Name:
豬褪黑素(MT)ELISA 試劑盒This kit can be used for determination of serum, plasma and liquid samples Organization Content.
The experimental principle:
The product levels were measured in samples of the kit by double antibody sandwichmethod. The product with the purified antibody coated microtiter plate, made of solid phase antibody, to package is the product antigen monoclonal antibodies are then added to the micropores, the product and then with HRP labeled antibody binding, the formation of antibody - antigen - antibody complex enzyme label, after thorough washing with TMB chromogenic substrate. TMB in the HRP enzyme catalytic conversion into the blue, and in the action of acid into the final yellow. This product is positively related to the depth of color and in the samples. Instrument measured absorbance in the 450nm wavelength with ELISA (OD), the product concentration in the samples was calculated by standard curve.
Materials provided with the kit
Materials provided with the kit | 48determinations | 96 determinations | Storage |
User manual | 1 | 1 |
|
Closure plate membrane | 2 | 2 |
|
Sealed bags | 1 | 1 |
|
Microelisa stripplate | 1 | 1 | 2-8℃ |
Standard:360ng/L | 0.5ml×1 bottle | 0.5ml×1 bottle | 2-8℃ |
Standard diluent | 1.5ml×1 bottle | 1.5ml×1 bottle | 2-8℃ |
HRP-Conjugate reagent | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
Sample diluent | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
Chromogen Solution A | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
Chromogen Solution B | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
Stop Solution | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
wash solution | (20ml×20 fold) ×1bottle | (20ml×30 fold) ×1bottle | 2-8℃ |
Specimen requirements
Assay procedure
1.Dilute and add sample to Standard: set 10 Standard wells on the ELISA plates coated, add Standard 100μl to the first and the second well, then add Standard dilution 50μl to the first and the second well, mix; take out 100μl form the first and the second well then add it to the third and the forth well separay. then add Standard dilution 50μl to the third and the forth well ,mix ; then take out 50μl from the third and the forth well discard, add 50μl to the fifth and the sixth well ,then add Standard dilution 50μl to the fifth and the sixth well, mix ; take out 50μl from the fifth and the sixth well and add to the seventh and the eighth well, then add Standard dilution 50μl to the seventh and the eighth well ,mix ; take out 50μl from the seventh and the eighth well and add to the ninth and the tenth well, add Standard dilution 50μl to the ninth and the tenth well, mix , take out 50μl from the ninth and the tenth well discard(add Sample 50μl to each well after Diluting ,(density: 240ng/L,160ng/L ,80ng/L,40ng/L, 20ng/L)
2.add sample:Set blank wells separay (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 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 by pat.
6.add enzyme:Add HRP-Conjugate reagent 50μl to each well, except blank well.
7.incubate:Operation with 3.
8.washing:Operation with 5.
9.color:Add Chromogen Solution A 50ul and Chromogen Solution B to each well, evade the light preservation 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).
11.assay:take blank well as zero , Read absorbance at 450nm after Adding Stop Solution and within 15min.
Important notes
Calculate:
Take the standard density as the horizontal, the OD value for the vertical ,draw the standard curve on graph paper, Find out the corresponding density according to the sample OD value by the Sample curve, multiplied by the dilution multiple, or calculate the straight line regression equation of the standard curve with the standard density and the OD value ,with the sample OD value in the equation, calculate the sample density, multiplied by the dilution factor, the result is the sample actual density.
豬褪黑素(MT)ELISA 試劑盒Storage and validity
1.Storage: 2-8℃.
2.validity: six months.
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