檢測標(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

檢測方法：ELISA

檢測類型：酶聯(lián)免疫夾心法

產(chǎn)品的用途：僅供科研究課題使用

價格及詳細(xì)資料：齊一生物銷售：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,價格公道,傾力為國內(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,我司將竭誠為您服務(wù)！本試劑盒用于測定血清,血漿及相關(guān)液體樣本中含量.

【人膠原凝集素（Collectin）ELISA試劑盒】實(shí)驗(yàn)原理：

   本試劑盒應(yīng)用雙抗體夾心法測定標(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波長下測定吸光度（OD值）,通過標(biāo)準(zhǔn)曲線計算樣品中該產(chǎn)品濃度.

試劑盒組成：

【人膠原凝集素（Collectin）ELISA試劑盒】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．不能檢測含NaN3的樣品,因NaN3抑制辣根過氧化物酶的（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）.

2.     加樣：分別設(shè)空白孔（空白對照孔不加樣品及酶標(biāo)試劑,其余各步操作相同）、待測樣品孔.在酶標(biāo)包被板上待測樣品孔中先加樣品稀釋液40μl,然后再加待測樣品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)（此時藍(lán)色立轉(zhuǎn)黃色）.

11. 測定：以空白空調(diào)零,450nm波長依序測量各孔的吸光度（OD值）. 測定應(yīng)在加終止液后15分鐘以內(nèi)進(jìn)行.

【人膠原凝集素（Collectin）ELISA試劑盒】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． 濃洗滌液可能會有結(jié)晶析出,稀釋時可在水浴中加溫助溶,洗滌時不影響結(jié)果.

3． 各步加樣均應(yīng)使用加樣器,并經(jīng)常校對其準(zhǔn)確性,以避免試驗(yàn)誤差.一次加樣時間控制在5分鐘內(nèi),如標(biāo)本數(shù)量多,推薦使用排搶加樣.

4． 請每次測定的同時做標(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）,請先用樣品稀釋液稀釋一定倍數(shù)（n倍）后再測定,計算時請*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).

【人膠原凝集素（Collectin）ELISA試劑盒】計算：

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,根據(jù)樣品的OD     

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

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

準(zhǔn)曲線的直線回歸方程式,將樣品的OD值     

代入方程式,計算出樣品濃度,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個月

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