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想要分別萃取出位在粒線體、細胞質與細胞核的蛋白質，觀察蛋白質在細胞內分布位置的變化嗎？ Abcam 所研發的 Cell Fractionation Kit - Standard (ab109719) 可以讓您簡便快速地從細胞樣本中萃取出分別位在粒線體 (mitochondria)、細胞質 (cytoplasm) 與細胞核 (nuclear) 的蛋白質，完全不需使用到任何機械性外力，僅需簡單離心步驟就能完成！ 本產品特別適合有需要觀察蛋白質在細胞內分布位置變化的科研人員使用。您可以在完成萃取步驟後直接使用 Western Blot (WB) 或 ELISA 檢測萃取產物的蛋白質表現量變化，例如在圖 1 的 WB 實驗結果中就可以觀察到 Jurkat 細胞、HeLa 細胞與 143B 細胞在加藥誘發產生細胞凋亡 (apoptosis) 後，細胞色素 c (cytochrome c) 的分布位置就從原本的粒線體 (M fraction) 轉移到細胞質 (C fraction)。
Figure 1. Apoptosis was induced in Jurkat, HeLa and 143B osteosarcoma cells by treatment with staurosporine or in Jurkat cells by FAS. Mitochondrial and cytoplasmic fractions were isolated using Cell Fractionation Kit - Standard (ab109719) and probed using Cytochrome c Apoptosis WB Antibody Cocktail (ab110415). As is clear from the gels, cytochrome c has translocated partially in FAS-induced cells and STS-treated osteosarcoma cells, and almost completely in STS-treated Jurkat and HeLa cells. The three control targets allow for verification of the "cleanness" of the cell fractionation. 圖 2 則是 HepG2 細胞經過 Cell Fractionation Kit - Standard (ab109719) 萃取分離後，以 WB 實驗檢測各種指標性蛋白質以確認分離效果。所檢測的蛋白質包含作為細胞質指標的 GAPDH (glyceraldehyde-3-phosphate dehydrogenase)、作為粒線體指標的 Hsp70、F1-ATPase α、PDH E1α (pyruvate dehydrogenase subunit E1α)、Cytochrome c，以及作為細胞核指標的 PARP (poly(ADP-ribose) polymerase) 與 SP1。
Figure 2. Cytoplasmic (C), mitchondrial (M) and nuclear (N) fractions of HepG2 cells were prepared as described in the protocol of Cell Fractionation Kit - Standard (ab109719). Fractions were analyzed by Western blotting using Cytochrome c Apoptosis WB Antibody Cocktail (ab110415) containing antibodies against mitochondrial matrix (pyruvate dehydrogenase subunit E1α, PDH E1α), mitochondrial inner membrane (F1-ATPase α (CV α)), mitochondrial intermembrane space (cytochrome c) and cytosolic (glyceraldehyde-3-phosphate dehydrogenase, GAPDH) markers as well as with antibodies against additional mitochondrial matrix (Hsp70) and nuclear (poly(ADP-ribose) polymerase, PARP and SP1) markers, followed by appropriate HRP-conjugated goat secondary antibodies and ECL detection. 若您有興趣進一步了解細胞各分部構造與胞器的指標性蛋白質專一性抗體產品，歡迎參考〈Subcellular and cell membrane markers - Antibodies against specific organelles, the cell membrane, or cytoskeletal components〉。完整產品資訊與最新活動訊息歡迎洽詢 Abcam 台灣代理 — 伯森生技。 Abcam 蛋白質萃取實驗產品訂購資訊
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