Sizing of protein A-colloidal gold probes for immunoelectron microscopy.
Sizing of protein A-colloidal gold probes for immunoelectron microscopy.
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DOI：
10.1083/jcb.90.2.533
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发表时间：
1981-08
影响因子：
7.8
通讯作者：
Geuze, H J
中科院分区：
文献类型：
--
作者：
Slot, J W;Geuze, H J
胶体溶液中的金颗粒通常在大小上变化很大。特别是最细的溶胶（直径小于15纳米）是非常不均匀的，方差系数（CV）为25- 35%。我们将葡萄球菌蛋白A与金颗粒（PA/Au）络合，然后用甘油或蔗糖梯度离心将其分离成非常均匀的亚组分。通过这种方法，可以制备4.5到15 nm之间几乎任何尺寸的PA/Au探针。这些组分中金颗粒的变化导致CV值在9 ~ 16%之间。PA/Au复合物的反应性不受梯度过程的影响，正如大鼠胰腺组织超薄切片的单标记和双标记免疫细胞化学所显示的那样。
Gold particles in colloidal solutions often vary considerably in size. The finest sols (diameter less than 15 nm), especially, are very heterogeneous, as is indicated by coefficients of variance (CV) of 25- 35%. We have complexed staphylococcal protein A with gold particles (PA/Au) and then fractionated the preparations by glycerol or sucrose gradient centrifugation into very homogeneous subfractions. In this way, PA/Au probes of almost any size between 4.5 and 15 nm could be prepared. The variation of the gold particles in these fractions resulted in CV's between 9 and 16%. The reactivity of the PA/Au complex was not affected by the gradient procedure, as was shown by single- and double-labeling immunocytochemistry of ultrathin cryosections of rat pancreatic tissue.