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人乙酰*酯酶(AChE)酶聯(lián)免疫(ELISA)試劑盒實(shí)驗(yàn)原理

閱讀:364發(fā)布時(shí)間:2013-05-27

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人乙酰*酯酶 (AChE)酶聯(lián)免疫分析(ELISA)試劑盒僅供研究使用,用于測定人血清,血漿,組織及相關(guān)液體樣本中乙酰*酯酶 (AChE)的含量。
實(shí)驗(yàn)原理:應(yīng)用雙抗體夾心法測定標(biāo)本中人乙酰*酯酶 (AChE)水平。用純化的人乙酰*酯酶 (AChE)抗體包被微孔板,制成固相抗體,往包被單抗的微孔中依次加入乙酰*酯酶 (ACh),再與HRP標(biāo)記的乙酰*酯酶 (AChE)抗體結(jié)合,形成抗體-抗原-酶標(biāo)抗體復(fù)合物,經(jīng)過*洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成zui終的黃色。顏色的深淺和樣品中的乙酰*酯酶 (AChE)呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(OD值),通過標(biāo)準(zhǔn)曲線計(jì)算樣品中人乙酰*酯酶 (AChE)濃度。
實(shí)驗(yàn)注意事項(xiàng):1.試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用,酶標(biāo)包被板開封后如未用完,板條應(yīng)裝入密封袋中保存。
2.濃洗滌液可能會(huì)有結(jié)晶析出,稀釋時(shí)可在水浴中加溫助溶,洗滌時(shí)不影響結(jié)果。
3.各步加樣均應(yīng)使用加樣器,并經(jīng)常校對(duì)其準(zhǔn)確性,以避免試驗(yàn)誤差。一次加樣時(shí)間控制在5分鐘內(nèi),如標(biāo)本數(shù)量多,推薦使用排槍加樣。
4.請(qǐng)每次測定的同時(shí)做標(biāo)準(zhǔn)曲線,做復(fù)孔。如標(biāo)本中待測物質(zhì)含量過高(樣本OD值大于標(biāo)準(zhǔn)品孔*孔的OD值),請(qǐng)先用樣品稀釋液稀釋一定倍數(shù)(n倍)后再測定,計(jì)算時(shí)請(qǐng)zui后乘以總稀釋倍數(shù)(×n×5)。
5.封板膜只限一次性使用,以避免交叉污染。
6.底物請(qǐng)避光保存。
7.嚴(yán)格按照說明書的操作進(jìn)行,試驗(yàn)結(jié)果判定必須以酶標(biāo)儀讀數(shù)為準(zhǔn).
8.所有樣品,洗滌液和各種廢棄物都應(yīng)按傳染物處理。
9.本試劑不同批號(hào)組分不得混用。
10. 如與英文說明書有異,以英文說明書為準(zhǔn)。
試劑盒保存:2-8℃。
有效期:6個(gè)月
Human Acetylcholinesterase
FOR RESEARCH USE ONLY
Drug Names
Generic Name:Human Acetylcholinesterase (AchE) ELISA Kit.
Purpose
This kit allows for the determination of AchE concentrations in Human serum, blood plasma, tissue and other biological fluids.
Principle of the assay
The kit assay Human AchE level in the sample,use Purified Human AchE antibody to coat microtiter plate wells, make solid-phase antibody, then add AchE to wells, Combined AchE antibody which With HRP labeled , 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 AchE in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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 mins, 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 diluente and 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:2-8℃.
validity:six months.
上海華壹生物科技有限公司
Shanghai HuaYi BioTechnology Co.,Ltd

:021-2420 9195,136 616 74336

【備注】:僅供科學(xué)研究實(shí)驗(yàn)使用,以上信息僅供參考,具體產(chǎn)品信息以隨貨說明書為準(zhǔn)。


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