Department of Congenital Diseases, Statens Serum Institut, Copenhagen S, Denmark.
Department of Autoimmunology and Biomarkers, Statens Serum Institut, Copenhagen, Denmark.
Methods Mol Biol. 2024;2821:249-263. doi: 10.1007/978-1-0716-3914-6_20.
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/absorption 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.
肽抗体在免疫细胞化学（ICC）和免疫组织化学（IHC）中特别有用，因为组织和细胞的固定可能导致抗原变性。肽抗体可以针对任何定义的序列进行制备，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）的可用性允许进行抑制/吸收对照，这在 ICC/IHC 中非常重要，因为免疫球蛋白 Fc 受体、非特异性反应以及一抗和二抗与其他抗原和内源性免疫球蛋白的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，并提出了适当对照的建议。
