Specificity controls for immunocytochemical methods
Specificity controls for immunocytochemical methods
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DOI：
10.1177/002215540004800201
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发表时间：
2000-02-01
影响因子：
3.2
通讯作者：
Burry, RW
中科院分区：
文献类型：
--
作者：
Burry, RW
免疫细胞化学用于细胞和组织中蛋白质的抗体定位。结果的特异性取决于两个独立的标准：抗体的特异性和所用方法的特异性。抗体特异性最好通过免疫印迹和/或免疫沉淀来确定。抗体与蛋白质的吸收并不能确定抗体是否会与组织中的相同蛋白质结合，因此不能很好地控制抗体特异性。该方法的特异性最好通过阴性对照（用血清代替一抗）和阳性对照（使用已知含有蛋白质的细胞的抗体）来确定。随着免疫细胞化学的使用越来越多，了解显示标记特异性所需的适当对照非常重要。
Immunocytochemistry is used for antibody localization of proteins in cells and tissues. The specificity of the results depends on two independent criteria: the specificity of the antibody and of the method used. The antibody specificity is best determined by immunoblot and or immunoprecipitation. Absorption of the antibody with a protein does not determine that the antibody would have bound to the same protein in the tissue, and therefore is not a good control for antibody specificity. The specificity of the method is best determined by both a negative control, replacing the primary antibody with serum, and a positive control, using the antibody with cells known to contain the protein. With the increasing use of immunocytochemistry, it is important to be aware of the appropriate controls needed to show specificity of the labeling.