Bio-Imaging Center, Rudolf Virchow Center, University of Würzburg, Würzburg, Germany.
Biotechniques. 2010 Aug;49(2):574-9. doi: 10.2144/000113466.
Here we describe a labeling technique for the covalent linkage of quantum dots to transmembrane receptors for single-molecule tracking. Our method combines the acyl carrier protein (ACP) technique with coenzyme A (CoA)-functionalized quantum dots to covalently attach quantum dots to ACP fusions of receptor proteins. The advantages of this approach include: (i) the use of a smaller attachment linker than in many other quantum dot-labeling systems; (ii) the ability to achieve a reliable 1:1 fluorophore-to-receptor labeling stoichiometry; (iii) the specificity of the method; and (iv) the covalent nature of the quantum dot linkage. We demonstrate the general suitability of this technique in single-molecule tracking, internalization, and trafficking studies by imaging two different transmembrane receptors in living cells.
在这里，我们描述了一种将量子点共价连接到跨膜受体的标记技术，用于单分子跟踪。我们的方法将酰基辅酶 A（ACP）技术与辅酶 A（CoA）功能化的量子点相结合，将量子点共价连接到受体蛋白的 ACP 融合物上。这种方法的优点包括：（i）使用比许多其他量子点标记系统更小的连接接头；（ii）能够实现可靠的 1:1 荧光团与受体标记物的比例；（iii）方法的特异性；以及（iv）量子点连接的共价性质。我们通过在活细胞中成像两种不同的跨膜受体，证明了该技术在单分子跟踪、内化和运输研究中的通用性。
