hce_kmu
105年
普通生物及生化概論
第 45 題
Which of the following sequence of the steps is regular for Western Blotting Assay that is widely used for detection of the specific protein in a tissue?
1. Tissues are broken down using a homogenizer or by sonication.
2. The protein is probed with an addition of the primary antibody.
3. The protein is transferred to a PVDF membrane.
4. Add secondary antibody conjugated with peroxidase.
5. The proteins of the tissues are separated using gel electrophoresis.
6. The protein is detected with an addition of a peroxidase substrate.
1. Tissues are broken down using a homogenizer or by sonication.
2. The protein is probed with an addition of the primary antibody.
3. The protein is transferred to a PVDF membrane.
4. Add secondary antibody conjugated with peroxidase.
5. The proteins of the tissues are separated using gel electrophoresis.
6. The protein is detected with an addition of a peroxidase substrate.
- A 123456
- B 153246
- C 152346
- D 154623
- E 124653
思路引導 VIP
想像你要在一大群混亂的馬拉松選手中找到一位穿特定號碼牌的人，但現場非常擁擠。你會選擇先讓選手們在跑道上按速度排開，還是直接在起點的混亂人群中尋找？另外，如果跑道（膠體）非常容易碎裂且厚重，不利於你貼上標籤，你會如何將這些排好隊的選手「拓印」到一張更薄、更好操作的定位紙上呢？
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AI 詳解
AI 專屬家教
恭喜你精確地掌握了西方墨點法（Western Blotting）的核心流程！這是一道檢驗生物技術基礎功的經典題目，你能從多個步驟中理出正確的邏輯，說明你對蛋白質分析的動態過程有很清晰的圖像。
蛋白質偵測的邏輯層次
整個實驗的邏輯是從「破壞」到「排序」，再到「偵測」。首先，我們必須透過**均質化（1）將組織破碎以釋放蛋白質；接著利用 SDS-PAGE 電泳（5），根據蛋白質分子的質量大小進行物理分離。由於膠體（Gel）質地脆弱且不便後續處理，關鍵的一步在於將蛋白質轉漬（3）到穩定的 PVDF 膜上。一旦目標物被固定在膜表面，偵測系統便能上場：先由一級抗體（2）特異性識別目標，再加入帶有酵素的二級抗體（4）進行訊號放大，最後投入受質（6）**產生呈色或發光訊號。
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