手機(jī)版
化工儀器網(wǎng)手機(jī)版
化工儀器網(wǎng)小程序
官方微信
公眾號(hào)：chem17
掃碼關(guān)注視頻號(hào)
Purpose
This kit allows for the determination oflipoteichoic acids（LTA）concentrations in Human serum, cellculture supernates and other biological fluids
Principle of the assay
The kit assay Human lipoteichoic acids（LTA）level in the sample，use Purified Human lipoteichoic acids（LTA）antibody to coat microtiter plate wells, make solid-phase antibody, then add lipoteichoic acids（LTA）to wells,Combined antibody which With HRP labeled goat anti- Human become antibody - antigen - enzyme-antibody complex, after washing Compley,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 Human lipoteichoic acids（LTA） 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 | Stopp Solution | 6ml×1 bottle |
| 2 | HRP-Conjugate reagent | 6ml×1 bottle | 8 | Standard（960pg/ml） | 0.5ml×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 requirements
1. extractas 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 preserve, Avoid repeated freeze-thaw cycles.
2. Can’t detect the sample which contain NaN3, because NaN3 inhibits HRP active.
Assay procedure
1. Dilute and add sample:Dilute Original density Standard as follow table:
| 480pg/ml | 5 Standard | 150μl Original density Standard+150μl Standard diluent |
| 240 pg/ml | 4 Standard | 150μl 5 Standard+150μl Standard diluent |
| 120 pg/ml | 3 Standard | 150μl 4 Standard+150μl Standard diluent |
| 60 pg/ml | 2 Standard | 150μl 3 Standard +150μl Standard diluent |
| 30 pg/ml | 1 Standard | 150μl 2 Standard +150μl Standard diluent |
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 wash solution diluted 30-fold （or 20-fold） with distilled water and reserve.
5.washing：Uncover Closure plate membrane, discardLiquid, 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.
Steps description
| Standard, Sample diluent |
| AddStandard, Sample diluent, incubate for 30 min at 37℃. |
| Wash 5 time,AddHRP-Conjugate reagent, incubate for 30 min at 37℃. |
| Wash 5 times,Add Chromogen Solution A and B, incubate for 30 min at 37℃. |
| AddStopp Solution |
| Read absorbance at 450nm within 15 min |
| calculate |
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.
Important notes
1. The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature, ELISA plates coated if has not use up after opened, the plate should be stored in Sealed bag.
2. washing buffer will Crystallization separation, it can be heated the water helps 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 of sample is much , recommend to use Volley .
4. if the testing material content is excessively higher （The sample OD is bigger than the first standard well ）,please dilute Sample （n-fold）, Please diluenteand multiplied by the dilution factor.（×n×5）.
5. Closure plate membrane only limits the disposable use, to avoid cross-contamination.
6. The substrate evade the light preservation.
7. Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a standard.
8. All samples, washing buffer and each kind of reject should according to infective material process.
9. Do not mix reagents with those from other lots.
Storage and validity
1．Storage： 2-8℃.
2．validity： six months.
相關(guān)產(chǎn)品
免責(zé)聲明
- 凡本網(wǎng)注明“來(lái)源：化工儀器網(wǎng)”的所有作品，均為浙江興旺寶明通網(wǎng)絡(luò)有限公司-化工儀器網(wǎng)合法擁有版權(quán)或有權(quán)使用的作品，未經(jīng)本網(wǎng)授權(quán)不得轉(zhuǎn)載、摘編或利用其它方式使用上述作品。已經(jīng)本網(wǎng)授權(quán)使用作品的，應(yīng)在授權(quán)范圍內(nèi)使用，并注明“來(lái)源：化工儀器網(wǎng)”。違反上述聲明者，本網(wǎng)將追究其相關(guān)法律責(zé)任。
- 本網(wǎng)轉(zhuǎn)載并注明自其他來(lái)源（非化工儀器網(wǎng)）的作品，目的在于傳遞更多信息，并不代表本網(wǎng)贊同其觀點(diǎn)和對(duì)其真實(shí)性負(fù)責(zé)，不承擔(dān)此類作品侵權(quán)行為的直接責(zé)任及連帶責(zé)任。其他媒體、網(wǎng)站或個(gè)人從本網(wǎng)轉(zhuǎn)載時(shí)，必須保留本網(wǎng)注明的作品第一來(lái)源，并自負(fù)版權(quán)等法律責(zé)任。
- 如涉及作品內(nèi)容、版權(quán)等問(wèn)題，請(qǐng)?jiān)谧髌钒l(fā)表之日起一周內(nèi)與本網(wǎng)聯(lián)系，否則視為放棄相關(guān)權(quán)利。
采購(gòu)中心