Background: Antibody validation for tissue staining is required for reproducibility; criteria to ensure validity have been published recently. The majority of these recommendations imply the use of routinely processed (formalin-fixed, paraffin-embedded) tissue. Materials & methods: We applied to lightly fixed frozen sections a panel of 126 antibodies validated for formalin-fixed, paraffin-embedded tissue with extended criteria. Results: Less than 30% of the antibodies performed as expected with all fixations. 35% preferred one fixation over another, 13% gave nonspecific staining and 23% did not stain at all. Conclusion: Individual antibody variability of the paratope's fitness for the fixed antigen may be the cause. Revalidation of established antibody panels is required when they are applied to sections whose fixation and processing are different from the tissue where they were initially validated.

中文翻译：

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经验证可用于常规处理组织的抗体对冰冻切片的染色不可预测。

背景：为了重现性，需要对组织染色进行抗体验证；最近公布了确保有效性的标准。这些建议中的大多数暗示使用常规处理（福尔马林固定、石蜡包埋）的组织。材料和方法：我们将一组 126 种抗体应用于轻度固定的冰冻切片，并根据扩展标准对福尔马林固定、石蜡包埋的组织进行了验证。结果：不到 30% 的抗体按预期执行所有固定。35% 的人更喜欢一种固定方式，13% 的人进行非特异性染色，23% 的人根本没有染色。结论：互补位对固定抗原的适应性的个体抗体变异性可能是原因。当将已建立的抗体板应用于固定和处理不同于它们最初验证的组织的切片时，需要对其进行重新验证。