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大鼠TNF-α ELISA試劑盒實驗原理及操作事項

時間:2011/10/13閱讀:1997
分享:
大鼠TNF-α  ELISA試劑盒使用目的:本試劑盒用于測定大鼠血清、血漿及相關液體樣本中TNF-α含量。實驗原理大鼠TNF-α  ELISA試劑盒應用雙抗體夾心法測定標本中大鼠TNF-α水平。用純化的大鼠TNF-α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TNF-α,再與HRP標記的TNF-α抗體結合,形成抗體-抗原-酶標抗體復合物,經過*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zui終的黃色。顏色的深淺和樣品中的TNF-α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大鼠TNF-α濃度。大鼠TNF-α  ELISA試劑盒注意事項1.試劑盒從冷藏環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。3.各步加樣均應使用加樣器,并經常校對其準確性,以避免試驗誤差。一次加樣時間控制在5分鐘內,如標本數量多,推薦使用排槍加樣。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(n倍)后再測定,計算時請zui后乘以總稀釋倍數(×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。

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