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</div><div>　　為確保實驗結(jié)果的準確性，正確使用<a href="http://m.ouke-automation.com/Products-37030636.html" target="_blank">大鼠低氧誘導因子1αelisa試劑盒</a>至關重要。以下是詳細的使用步驟：</div><div> </div><p align="center"><img src="https://img42.chem17.com/9/20250211/638748686118437150289.jpg" width="500" height="500" align="middle" alt="" /></p><p> </p><div>　　一、實驗前準備</div><div> </div><div>　　1、試劑盒檢查：確認試劑盒包含標準品、檢測抗體、酶結(jié)合物、底物、終止液、洗滌液等，并檢查是否在有效期內(nèi)。</div><div> </div><div>　　2、樣本準備：采集大鼠組織或細胞樣本，按要求處理（如勻漿、離心）并保存于-80°C。</div><div> </div><div>　　3、試劑平衡：將試劑盒從冰箱取出，室溫平衡30分鐘。</div><div> </div><div>　　4、儀器準備：確保酶標儀、移液器、離心機等設備正常工作。</div><div> </div><div>　　二、實驗步驟</div><div> </div><div>　　1、標準品稀釋：按說明書將標準品梯度稀釋，通常設置6-8個濃度點。</div><div> </div><div>　　2、加樣：將標準品和樣本加入酶標板孔中，每孔100μL，設置空白對照。</div><div> </div><div>　　3、孵育：37°C孵育1-2小時，使HIF-1α與包被抗體結(jié)合。</div><div> </div><div>　　4、洗滌：棄去孔內(nèi)液體，加入洗滌液，靜置30秒后棄去，重復3-5次。</div><div> </div><div>　　5、加檢測抗體：每孔加入100μL生物素標記的檢測抗體，37°C孵育1小時。</div><div> </div><div>　　6、洗滌：重復洗滌步驟。</div><div> </div><div>　　7、加酶結(jié)合物：每孔加入100μL酶結(jié)合物，37°C孵育30分鐘。</div><div> </div><div>　　8、洗滌：再次洗滌。</div><div> </div><div>　　9、加底物：每孔加入100μL底物溶液，避光孵育15-30分鐘。</div><div> </div><div>　　10、終止反應：每孔加入50μL終止液，顏色由藍變黃。</div><div> </div><div>　　11、讀數(shù)：用酶標儀在450nm波長下測定吸光度。</div><div> </div><div>　　三、數(shù)據(jù)分析</div><div> </div><div>　　1、繪制標準曲線：以標準品濃度為橫坐標，吸光度為縱坐標，繪制標準曲線。</div><div> </div><div>　　2、計算樣本濃度：根據(jù)標準曲線計算樣本中HIF-1α的濃度。</div><div> </div><div>　　四、注意事項</div><div> </div><div>　　1、操作規(guī)范：避免污染，使用一次性吸頭。</div><div> </div><div>　　2、時間控制：嚴格控制孵育和洗滌時間。</div><div> </div><div>　　3、儀器校準：確保酶標儀和移液器準確。</div><div> </div><div>　　4、重復實驗：每個樣本至少做3次重復，確保結(jié)果可靠。</div> </ul> <div class="tjnxtzn5ak" id="news_sx"> <li id="tjnxtzn5ak" class="sx_left">上一篇：沒有了<br />下一篇：<a href="/Article-3923716.html">一文與您分享動物ELISA試劑盒的常見問題相應解決方法</a></li> <li id="tjnxtzn5ak" class="sx_right"><a href="/article.html">返回</a></li> </div> </div> </div> </div> </div> <div class="tjnxtzn5ak" id="clear"></div> <div id="tjnxtzn5ak" class="footer"> 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