Antibodies validated for routinely processed tissue unpredictably stain frozen tissue sections.

Background: Antibody validation for tissue staining is required for reproducibility and criteria to ensure validity have been published recently. The majority of these recommendations imply the use of routinely processed tissue (FFPE).
Materials & Methods: We applied to lightly fixed frozen sections a panel of 126 antibodies validated for FFPE with extended criteria.
Results: Less than 30% performed conservatively with all fixations, 35% preferred one fixation over another, 13% gave non-specific staining, 23% did not stain at all.
Conclusions: Individual antibody variability of the paratope fitness for the fixed antigen may be the cause. Re-validation of established antibody panels is required when applied to sections whose fixation and processing is different from the tissue where they have been initially validated.
These are supplementary Data to the manuscript. In addition Tabel 1 and Supplementary Table 1 are provided in Excel format.

背景：组织染色所用抗体的验证工作是保障研究可重复性的必要前提，近期已有保障抗体有效性的相关标准正式发表。此类推荐方案的绝大多数均提及需使用常规处理的组织样本，即福尔马林固定石蜡包埋（FFPE）样本。
材料与方法：本研究将126种经扩展验证标准确证可用于福尔马林固定石蜡包埋（FFPE）样本的抗体组，应用于轻度固定的冰冻切片。
结果：仅不到30%的抗体在所有固定方式下均表现稳定；35%的抗体仅适配某一种固定方式；13%的抗体出现非特异性染色；另有23%的抗体完全无法显色。
结论：抗体互补位适配固定化抗原的能力存在个体差异，或为上述实验现象的成因。当已验证的抗体组应用于固定方式与处理流程均与初始验证样本不同的切片时，需对其重新进行验证。
本内容为该研究论文的补充数据。此外，表1与补充表1均以Excel格式提供。