小鼠肝素結合性表皮生長因子(HB-EGF)ELISA試劑盒性能:
1. 靈敏度:Z小的檢測濃度小于1號標準品。稀釋度的線性。樣品線性回歸與預期濃度相關系數R值為0.990。
2. 特異性:不與其它細胞因子反應。
3. 重復性:板內、板間變異系數均小于10%。
小鼠肝素結合性表皮生長因子(HB-EGF)ELISA試劑盒
英文名稱:Mouse heparin-binding epidermal growth factor-like growth factor,HB-EGF Elisa Kit
貨號:K-M4920
所需樣本體積: 50-100ul
檢測波長: 450 nm
用途: For research use only. Not for diagnostic use.
小鼠肝素結合性表皮生長因子(HB-EGF)ELISA試劑盒操作步驟:
1. 使用前,將所有試劑充分混勻。不要使液體產生大量的泡沫,以免加樣時加入大量的氣泡,產生加樣上的誤差。
2. 根據待測樣品數量加上標準品的數量決定所需的板條數。每個標準品和空白孔建議做復孔。每個樣品根據自己的數量來定,能使用復孔的盡量做復孔。
3. 加入稀釋好后的標準品50ul于反應孔、加入待測樣品50ul于反應孔內。立即加入50ul的生物素標記的抗體。蓋上膜板,輕輕振蕩混勻,37℃溫育45分鐘。
4. 甩去孔內液體,每孔加滿洗滌液,振蕩30秒,甩去洗滌液,用吸水紙拍干。重復此操作4次。如果用洗板機洗滌,洗滌次數增加一次。
5. 每孔加入100ul的親和鏈酶素-HRP,輕輕振蕩混勻,37℃溫育30分鐘。
6. 甩去孔內液體,每孔加滿洗滌液,振蕩30秒,甩去洗滌液,用吸水紙拍干。重復此操作4次。如果用洗板機洗滌,洗滌次數增加一次。
7. 每孔加入底物A、B各50ul,輕輕振蕩混勻,37℃溫育5分鐘。避免光照。
8. 取出酶標板,迅速加入50ul終止液,加入終止液后應立即測定結果。
9. 在450nm波長處測定各孔的OD值。
結果判斷與分析:
1、 儀器值:于波長450nm的酶標儀上讀取各孔的OD值
2、 以吸光度OD值為縱坐標(Y),相應的LTB4標準品濃度為橫坐標(X),做得相應的曲線,樣品的LTB4含量可根據其OD值由標準曲線換算出相應的濃度,再乘以稀釋倍數;或用標準物的濃度與OD值計算出標準曲線的回歸方程式,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,即為樣品的實際濃度。
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