Friis Tina, Pedersen Klaus Boberg, Hougaard David, Houen Gunnar
Department of Autoimmunology and Biomarkers, Statens Serum Institut, Artillerivej 5, 2300, Copenhagen, Denmark.
Department of Congenital Diseases, Statens Serum Institut, Copenhagen, Denmark.
Methods Mol Biol. 2015;1348:311-25. doi: 10.1007/978-1-4939-2999-3_27.
Peptide antibodies are particularly useful for immunocytochemistry (ICC) and immunohistochemistry (IHC), where antigens may denature due to fixation of tissues and cells. Peptide antibodies can be made to any defined sequence, including unknown putative proteins and posttranslationally modified sequences. Moreover, the availability of large amounts of the antigen (peptide) allows inhibition/adsorption controls, which are important in ICC/IHC, due to the many possibilities for false-positive reactions caused by immunoglobulin Fc receptors, nonspecific reactions, and cross-reactivity of primary and secondary antibodies with other antigens and endogenous immunoglobulins, respectively. Here, simple protocols for ICC and IHC are described together with recommendations for appropriate controls.
肽抗体对于免疫细胞化学（ICC）和免疫组织化学（IHC）尤为有用，在这些技术中，由于组织和细胞的固定，抗原可能会变性。可以针对任何确定的序列制备肽抗体，包括未知的假定蛋白质和翻译后修饰的序列。此外，大量抗原（肽）的可获得性使得抑制/吸附对照成为可能，这在ICC/IHC中很重要，因为免疫球蛋白Fc受体、非特异性反应以及一抗和二抗分别与其他抗原和内源性免疫球蛋白发生交叉反应会导致许多假阳性反应的可能性。本文描述了ICC和IHC的简单方案以及适当对照的建议。
