Optimization of a thermochemical antibody stripping method for enhanced tyramide signal amplification-based opal multiplex immunohistochemistry in fragile tissues
Evaluation of microwave oven-assisted antibody removal (MO-AR), chemical reagent-based antibody removal (CR-AR), and hybridization oven-based stripping at 50 °C (HO-AR-50) and 98 °C (HO-AR-98) revealed that MO-AR and HO-AR-98 most effectively removed primary and secondary antibodies. In five-color mIHC on mouse kidney sections, both methods yielded strong target-specific signals. However, in brain tissue sections prone to delamination, HO-AR-98 better preserved tissue integrity compared to MO-AR and supported effective multi-target staining. These findings establish a novel thermochemical stripping method compatible with TSA-based Opal mIHC, enhancing its utility in research and clinical diagnostics.
Authors
- Lixiang Xue (ORCID: https://orcid.org/0000-0003-1507-7193)
- Yan Song (ORCID: https://orcid.org/0000-0002-6729-2672)
- Chen Li
- Chen Huang (ORCID: https://orcid.org/0000-0002-2407-9716)
- Jianling Yang (ORCID: https://orcid.org/0000-0003-3260-4346)
- Xiaotong Yu (ORCID: https://orcid.org/0000-0002-0795-3846)
Institutions
- Peking University (CN)
- Peking University Third Hospital (CN)
Publication Details
- Journal
- BMC Research Notes
- Published
- 2025-05-27
- DOI
- https://doi.org/10.1186/s13104-025-07301-4
- Primary Topic
- Monoclonal and Polyclonal Antibodies Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Natural Science Foundation of China