檢測(cè)標(biāo)本：血清,血漿,尿液,胸腹水,腦脊液,細(xì)胞培養(yǎng)上清,組織勻漿等

檢測(cè)方法：ELISA

檢測(cè)類(lèi)型：酶聯(lián)免疫夾心法

產(chǎn)品的用途：僅供科研究課題使用

價(jià)格及詳細(xì)資料：齊一生物銷(xiāo)售：021－6034 8496；181214 53965；173021 04490；.齊一生物科技（上海）有限公司提供的ELISA試劑盒受到了廣大科研單位的一致肯定和認(rèn)同.大品牌保證,價(jià)格公道,傾力為國(guó)內(nèi)外科研院校實(shí)驗(yàn)室提供*優(yōu)質(zhì)的產(chǎn)品.若有需要,我司將竭誠(chéng)為您服務(wù)！本試劑盒用于測(cè)定血清,血漿及相關(guān)液體樣本中含量.

【人絲裂原活化蛋白激酶激酶6(MKK6)ELISA試劑盒廠家】實(shí)驗(yàn)原理：

   本試劑盒應(yīng)用雙抗體夾心法測(cè)定標(biāo)本中該產(chǎn)品水平.用純化的本產(chǎn)品抗體包被微孔板,制成固相抗體,往包被單抗的微孔中依次加入本產(chǎn)品抗原,再與HRP標(biāo)記的本產(chǎn)品抗體結(jié)合,形成抗體-抗原-酶標(biāo)抗體復(fù)合物,經(jīng)過(guò)徹底洗滌后加底物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*終的黃色.顏色的深淺和樣品中的本產(chǎn)品呈正相關(guān).用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度（OD值）,通過(guò)標(biāo)準(zhǔn)曲線(xiàn)計(jì)算樣品中該產(chǎn)品濃度.

試劑盒組成：

齊一生物銷(xiāo)售：021－6034 8496；181214 53965；173021 04490

【人絲裂原活化蛋白激酶激酶6(MKK6)ELISA試劑盒廠家】標(biāo)本要求：

1．標(biāo)本采集后盡早進(jìn)行提取,提取按相關(guān)文獻(xiàn)進(jìn)行,提取后應(yīng)盡快進(jìn)行實(shí)驗(yàn).若不能馬上進(jìn)行試驗(yàn),可將標(biāo)本放于-20℃保存,但應(yīng)避免反復(fù)凍融

2．不能檢測(cè)含NaN3的樣品,因NaN3抑制辣根過(guò)氧化物酶的（HRP）活性.

操作步驟：

1.     標(biāo)準(zhǔn)品的稀釋與加樣：在酶標(biāo)包被板上設(shè)標(biāo)準(zhǔn)品孔10孔,在、第二孔中分別加標(biāo)準(zhǔn)品100μl,然后在、第二孔中加標(biāo)準(zhǔn)品稀釋液50μl,混勻；然后從孔、第二孔中各取100μl分別加到第三孔和第四孔,再在第三、第四孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻；然后在第三孔和第四孔中先各取50μl棄掉,再各取50μl分別加到第五、第六孔中,再在第五、第六孔中分別加標(biāo)準(zhǔn)品稀釋液50ul,混勻；混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中,再在第七、第八孔中分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第七、第八孔中分別取50μl加到第九、第十孔中,再在第九第十孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第九第十孔中各取50μl棄掉.（稀釋后各孔加樣量都為50μl,濃度分別為1200 ng/L,800 ng/L ,400 ng/L,200ng/L, 100 ng/L）. 【人絲裂原活化蛋白激酶激酶6(MKK6)ELISA試劑盒廠家】

2.     加樣：分別設(shè)空白孔（空白對(duì)照孔不加樣品及酶標(biāo)試劑,其余各步操作相同）、待測(cè)樣品孔.在酶標(biāo)包被板上待測(cè)樣品孔中先加樣品稀釋液40μl,然后再加待測(cè)樣品10μl（樣品*終稀釋度為5倍）.加樣將樣品加于酶標(biāo)板孔底部,盡量不觸及孔壁,輕輕晃動(dòng)混勻.

3.     溫育：用封板膜封板后置37℃溫育30分鐘.

4.     配液：將30（48T的20倍）倍濃縮洗滌液用蒸餾水30（48T的20倍）倍稀釋后備用.

5.     洗滌：小心揭掉封板膜,棄去液體,甩干,每孔加滿(mǎn)洗滌液,靜置30秒后棄去,如此重復(fù)5次,拍干.

6.     加酶：每孔加入酶標(biāo)試劑50μl,空白孔除外.

7.     溫育：操作同3.

8.     洗滌：操作同5.

9.     顯色：每孔先加入顯色劑A50μl,再加入顯色劑B50μl,輕輕震蕩混勻,37℃避光顯色15分鐘.

10. 終止：每孔加終止液50μl,終止反應(yīng)（此時(shí)藍(lán)色立轉(zhuǎn)黃色）.

11. 測(cè)定：以空白空調(diào)零,450nm波長(zhǎng)依序測(cè)量各孔的吸光度（OD值）. 測(cè)定應(yīng)在加終止液后15分鐘以?xún)?nèi)進(jìn)行.

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(kāi)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. 【人絲裂原活化蛋白激酶激酶6(MKK6)ELISA試劑盒廠家】

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,推薦使用排搶加樣.

4． 請(qǐng)每次測(cè)定的同時(shí)做標(biāo)準(zhǔn)曲線(xiàn),做復(fù)孔.如標(biāo)本中待測(cè)物質(zhì)含量過(guò)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(cè)定,計(jì)算時(shí)請(qǐng)*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(shuō)明書(shū)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(shuō)明書(shū)有異,以英文說(shuō)明書(shū)為準(zhǔn).

計(jì)算：

以標(biāo)準(zhǔn)物的濃度為橫坐標(biāo),OD值為縱坐標(biāo),   

在坐標(biāo)紙上繪出標(biāo)準(zhǔn)曲線(xiàn),根據(jù)樣品的OD     

值由標(biāo)準(zhǔn)曲線(xiàn)查出相應(yīng)的濃度；再乘以稀釋     

倍數(shù)；或用標(biāo)準(zhǔn)物的濃度與OD值計(jì)算出標(biāo)     

準(zhǔn)曲線(xiàn)的直線(xiàn)回歸方程式,將樣品的OD值     

代入方程式,計(jì)算出樣品濃度,再乘以稀釋

倍數(shù),即為樣品的實(shí)際濃度.

試劑盒性能：

1.樣品線(xiàn)性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.95以上.

2.批內(nèi)與批見(jiàn)應(yīng)分別小于9%和11%    

保存條件及有效期：

1.試劑盒保存；2-8℃.

2．有效期：6個(gè)月

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990393     Reagent Bottles, 30 ml (6)                CNY                             

990392     Rotor Adapter Holder               CNY                             

940054     BioSprint 96 DNA Blood Kit (48)                CNY                             

939018     Deparaffinization solution (50ml)             CNY                             

331451     miScript PreAMP PCR Kit (12)           CNY                             

1070498  Buffer TCL, 2x (8.5 ml),G          CNY                             

1070211  Cramp (1)                   CNY                             

981313     Rotor-Disc 100 (300)                  CNY                             

981008     PCR Tubes, 0.2 ml  (10000)               CNY                             

9012945  Liquid Handling Kit Plato 3000 (500ul)              CNY                             

9012821  Liquid Handling Kit Plato 3301 (1000ul)            CNY                             

133257     NeXtalStock HEPES sod. salt pH 7.6 (200)                 CNY                             

330612     RT² SYBR Green Fluor FAST Mastermix (12)            CNY                              【人絲裂原活化蛋白激酶激酶6(MKK6)ELISA試劑盒廠家】

330521     RT² SYBR Green ROX qPCR Mastermix (8)                CNY                             

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206544     Type-it HRM PCR Kit (400)                 CNY                             

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9012737  Syringe pump outlet tubing (4.7m)           CNY                             

9012720  Set of Pipet Tubing Plato 3001                   CNY                             

9012717  Troughs, 50ml (100 troughs + lids)            CNY                             

133237     NeXtalStock Sodium citrate pH 5.4 (200)                  CNY                             

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