產(chǎn)品展示
人鈣衛(wèi)蛋白(calprotein)ELISA試劑盒
點擊次數(shù):14發(fā)布時間:2016/6/9 20:53:02

更新日期:2016/6/9 20:53:02
所 在 地:中國大陸
產(chǎn)品型號:
優(yōu)質供應
詳細內容
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,傾力為國內外科研院校實驗室提供*優(yōu)質的產(chǎn)品.若有需要,我司將竭誠為您服務!本試劑盒用于測定血清,血漿及相關液體樣本中含量.
【人鈣衛(wèi)蛋白(calprotein)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酶的催化下轉化成藍色,并在酸的作用下轉化成*終的黃色.顏色的深淺和樣品中的本產(chǎn)品呈正相關.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:
試劑盒組成 | 48孔配置 | 96孔配置 | 保存 |
說明書 | 1份 | 1份 |
|
封板膜 | 2片(48) | 2片(96) |
|
密封袋 | 1個 | 1個 |
|
酶標包被板 | 1×48 | 1×96 | 2-8℃保存 |
標準品:1800ng/L | 0.5ml×1瓶 | 0.5ml×1瓶 | 2-8℃保存 |
標準品稀釋液 | 1.5ml×1瓶 | 1.5ml×1瓶 | 2-8℃保存 |
酶標試劑 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 |
樣品稀釋液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 |
顯色劑A液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 |
顯色劑B液 | 3 ml×1瓶 | 6 ml×1瓶 | 2-8℃保存 |
終止液 | 3ml×1瓶 | 6ml×1瓶 | 2-8℃保存 |
濃縮洗滌液 | (20ml×20倍)×1瓶 | (20ml×30倍)×1瓶 | 2-8℃保存 |
齊一生物銷售:021-6034 8496;181214 53965;173021 04490
【人鈣衛(wèi)蛋白(calprotein)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). 【人鈣衛(wèi)蛋白(calprotein)ELISA試劑盒】
2. 加樣:分別設空白孔(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(此時藍色立轉黃色).
11. 測定:以空白空調零,450nm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晶析出,稀釋時可在水浴中加溫助溶,洗滌時不影響結果. 【人鈣衛(wèi)蛋白(calprotein)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,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系數(shù)R值為0.95以上.
2.批內與批見應分別小于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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