Jahn R, Schiebler W, Greengard P
Proc Natl Acad Sci U S A. 1984 Mar;81(6):1684-7. doi: 10.1073/pnas.81.6.1684.
An immunoassay method is described for the quantitative determination of synapsin I (protein I) and of a 36,000-dalton membrane protein from rat brain synaptic vesicles. The samples are spotted on nitrocellulose membrane filters, incubated sequentially with specific antibodies and 125I-labeled protein A, and assayed for radioactivity in a gamma scintillation counter. Conditions have been established to prevent losses of protein from the sheets during processing, to quench background radioactivity, and to adjust the sensitivity to the range desired. A large number of samples can be handled in parallel. The assay does not require iodination of the antigen and is accurate even with crude tissue samples. Standard curves were linear over a 20- to 50-fold range. The sensitivity of the method is such that 10 pmol of synapsin I and 50 ng of total vesicle membrane protein could be measured with accuracy. The method should prove useful for a wide range of proteins.
本文描述了一种免疫分析方法，用于定量测定大鼠脑突触小泡中的突触素I（蛋白I）和一种36,000道尔顿的膜蛋白。将样品点样在硝酸纤维素膜滤器上，依次与特异性抗体和125I标记的蛋白A孵育，然后在γ闪烁计数器中测定放射性。已经确定了相关条件，以防止在处理过程中蛋白质从滤膜上损失，淬灭背景放射性，并将灵敏度调整到所需范围。大量样品可以并行处理。该分析方法不需要对抗原进行碘化，即使对于粗制组织样品也很准确。标准曲线在20至50倍的范围内呈线性。该方法的灵敏度足以准确测定10皮摩尔的突触素I和50纳克的总囊泡膜蛋白。该方法对多种蛋白质应是有用的。
