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別藻藍蛋白 (APC)
抗體接合試劑套組
​關於別藻藍蛋白 (APC)
抗體接合試劑套組

別藻藍蛋白 (APC) 是由一種微藻所萃取純化而成,因藻類生長不易,為市面上較稀少的螢光染劑。以近紅外雷射光 (650 nm) 激發後,散射光波峰位在 (660 nm) 因此可與其他螢光染劑、檢體(如:細胞、蛋白)本身所發出的自體螢光做出區隔,增加檢測的便利度及判讀性,同時也能搭配其他染劑作多染觀測。別藻藍蛋白 (APC) 廣泛被使用於流式細胞儀、螢光顯微鏡等螢光檢測儀器。

 

鴻藻的別藻藍蛋白 (APC) 檢測套組為 SH-Reactive,操作者​無須再將其活化即可直接與抗體反應,完整操作及反應時間共 3 小時。

實驗操作範例 (SH-Reactive)
  1. 將反應溶液加入抗體中,置於室溫下反應 30 分鐘。

  2. 加入預先已活化完畢的 R-PE 並置於室溫下避光反應 1 小時。

  3. 將滅活劑加入混和溶液中,並置於室溫下避光反應30 分鐘。

  4. ​螢光蛋白抗體接合完成。

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  • Are phycobiliproteins stored in PBS only and in PBS with ammonium sulfate different?
    PBPs in different buffer solutions both exhibit good long-term stability, the form with ammonium sulfate requires more purification processes, such as dialysis or sulfate precipitation, to fully remove the ammonium sulfate prior to making tandem dyes and antibody conjugations. Home Algae Biotech offers phycobiliproteins stored in PBS only and PBS with ammonium sulfate buffer solutions.
  • Is the conjugation reaction linking the R-PE/APC to free sulfhydryl groups on the antibody?
    Home Algae's R-PE & APC Labeling Kit are SH-reactive labeling kits. Crosslinked R-PE and APC have maleimide (-NH2) groups that easily react with sulfhydryl (-SH) groups of target antibody without the need for additional activation. The maleinide group of R-PE/APC and sulfhydryl group on the antibody form a covalent bond during conjugation. Users can also use crosslinked R-PE & APC to label other molecules that have free sulfhydryl groups.
  • Do I need to remove the free R-PE from the R-PE-antibody conjugate?
    After the conjugation procedures, reaction mixture should contain mostly the R-PE-antibody conjugate, with small amounts of unconjugated R-PE and antibody. Unconjugated R-PE should not interfere with the assay result. However, purification is suggested for labeling up to 1 mg of antibody. Please contact us for more technical advice.
  • Can I purchase large pack of labeling kit and separate them into small vials on my own for several reactions?
    Yes, all pack sizes of our labeling kits are scalable and can be separated into small packs based on your experiment requirements.
  • What are the molecular weight of purified R-PE and APC?
    The molecular weight of purified R-PE is 240 kDa, and for purified APC is 105 kDa. They are both perfect for detecting cell surface antigens to further conduct cell screening and cell imaging. With high molecular weight, PE and APC won't penetrate into the cell sample even if cell membrane ruptured, which greatly lower the risk of false-positive reaction. It should be noted that the relatively high molecular weight of R-PE may preclude its use in procedures requiring good penetration into cells and tissues. It is predominantly intended for surface labeling of cells for flow cytometry.
​別藻藍蛋白 (APC) 優勢
  • 適合進行多種蛋白同時染色觀測

    • 由流式細胞儀、螢光顯微鏡雷射光 (650 nm) 即能激發。

    • 吸收光與散射光訊號間隔明顯​。​​

  • 確保檢測結果精確性

    • ​Brightness Index 列第四級,低衰退率、光穩定性佳,不易淬滅。

    • 分子量大,即便實驗過程細胞膜破裂,染劑也不會滲透⾄細胞內造成偽陽性。

  • 實驗操作安全

    • 純天然海洋⽣物萃取、無毒,不影響實驗樣本特性,對人體、血液檢體、環境皆無害。

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