SHANK3 Antibody Validation: Differential Performance in Western Blotting, Immunocyto- and Immunohistochemistry.

IF 2.8 4区医学 Q2 NEUROSCIENCES

Frontiers in Synaptic Neuroscience Pub Date : 2022-06-06 eCollection Date: 2022-01-01 DOI:10.3389/fnsyn.2022.890231

Anne-Kathrin Lutz, Helen Friedericke Bauer, Valentin Ioannidis, Michael Schön, Tobias M Boeckers

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引用次数: 8

Abstract

SHANK3 is a scaffolding protein implicated in autism spectrum disorders (ASD). Its function at excitatory glutamatergic synapses has been studied for the last two decades, however, tissue-specific expression patterns as well as its subcellular localization need to be studied in further detail. Especially the close sequence homology of SHANK3 to its protein family members SHANK2 and SHANK1 raises the emerging need for specific antibodies that are validated for the desired methodology. With this study, we aim to validate a set of commercial as well as homemade SHANK3 antibodies in Western Blotting, and synaptic immunocyto- and immunohistochemistry. We found that only a small subset of the antibodies included in this study meet the criteria of quality and specificity. Therefore, we aim to share our findings on SHANK3 antibody validation but also raise awareness of the necessity of antibody specificity testing in the field.

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SHANK3抗体验证:免疫印迹，免疫细胞和免疫组织化学的差异性能。

SHANK3是一种与自闭症谱系障碍(ASD)有关的支架蛋白。其在兴奋性谷氨酸突触中的功能已被研究了近二十年，然而，组织特异性表达模式及其亚细胞定位需要进一步详细研究。特别是SHANK3与其蛋白家族成员SHANK2和SHANK1的序列同源性引起了对特定抗体的需求，这些抗体需要经过所需方法的验证。在这项研究中，我们的目标是在Western Blotting、突触免疫细胞化学和免疫组织化学中验证一套商业和自制的SHANK3抗体。我们发现，本研究中只有一小部分抗体符合质量和特异性标准。因此，我们的目标是分享我们在SHANK3抗体验证方面的发现，同时也提高人们对该领域抗体特异性检测必要性的认识。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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