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

產(chǎn)品的用途：僅供科研究課題使用

價格及詳細資料：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同.大品牌保證,價格公道,傾力為國內(nèi)外科研院校實驗室提供*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.

【人蛋白酶3抗體（PR3Ab）ELISA試劑盒】實驗原理：

   本試劑盒應用雙抗體夾心法測定標本中該產(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標記的本產(chǎn)品抗體結(jié)合,形成抗體-抗原-酶標抗體復合物,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).用酶標儀在450nm波長下測定吸光度（OD值）,通過標準曲線計算樣品中該產(chǎn)品濃度.

試劑盒組成：

齊一生物銷售：021－6034 8496；181214 53965；173021 04490

【人蛋白酶3抗體（PR3Ab）ELISA試劑盒】標本要求：

1．標本采集后盡早進行提取,提取按相關(guān)文獻進行,提取后應盡快進行實驗.若不能馬上進行試驗,可將標本放于-20℃保存,但應避免反復凍融

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.     標準品的稀釋與加樣：在酶標包被板上設(shè)標準品孔10孔,在、第二孔中分別加標準品100μl,然后在、第二孔中加標準品稀釋液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、第四孔分別加標準品稀釋液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、第六孔中分別加標準品稀釋液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、第八孔中分別加標準品稀釋液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準品稀釋液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）. 【人蛋白酶3抗體（PR3Ab）ELISA試劑盒】

2.     加樣：分別設(shè)空白孔（空白對照孔不加樣品及酶標試劑,其余各步操作相同）、待測樣品孔.在酶標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5次,拍干.

6.     加酶：每孔加入酶標試劑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（此時藍色立轉(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）. 測定應在加終止液后15分鐘以內(nèi)進行.

注意事項：

1． 試劑盒從冷藏環(huán)境中取出應在室溫平衡15-30分鐘后方可使用,酶標包被板開封后如未用完,板條應裝入密封袋中保存.

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. 【人蛋白酶3抗體（PR3Ab）ELISA試劑盒】

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

4． 請每次測定的同時做標準曲線,做復孔.如標本中待測物質(zhì)含量過高（樣本OD值大于標準品孔孔的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． 嚴格按照說明書的操作進行,試驗結(jié)果判定必須以酶標儀讀數(shù)為準.

8． 所有樣品,洗滌液和各種廢棄物都應按傳染物處理.

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.

計算：

以標準物的濃度為橫坐標,OD值為縱坐標,   

在坐標紙上繪出標準曲線,根據(jù)樣品的OD     

值由標準曲線查出相應的濃度；再乘以稀釋     

倍數(shù)；或用標準物的濃度與OD值計算出標     

準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),即為樣品的實際濃度.

試劑盒性能：

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

2.批內(nèi)與批見應分別小于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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