技术文章

Analysis and handling of IHC common problems

Problem Reason Suggest
The result is too dark The concentration of the primary antibody is too high or the incubation time is too long Reduce primary antibody concentration or incubation time
The result is too dark The incubation temperature is too high Incubate at room temperature or 4 °C
The result is too dark The incubation time of the HRP-labeled secondary antibody is too long Reduce time
Non-specific staining in IHC results The paraffin section is not completely dewaxed Extend deparaffinization time
Non-specific staining in IHC results Insufficient rinsing during the procedure Increase the number and duration of washes
Non-specific staining in IHC results The tissue is rich in endogenous biotin and peroxidase Block with relevant reagent
Non-specific staining in IHC results Antigenic ectopia occurs
Non-specific staining in IHC results Insufficient protein blocking Increase the protein blocking time
Weak color development in IHC results The concentration of the primary antibody is too low or the incubation time is too short Increase the concentration of the primary antibody or extend the incubation time
Weak color development in IHC results The reagent has expired Confirm the shelf life of the reagent before the experiment
Weak color development in IHC results Excessive rinsing during the procedure Rinse appropriately
No staining in IHC results Expression of non-target antigens in the tissue
No staining in IHC results mismatch between the species of the primary antibody and the secondary antibody Verify the species origin of the primary antibody before the experiment
No staining in IHC results mismatch between the chromogen and the HRP system Verify the color development system before the experiment


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目录: 技术文章
探索主题: Immunohistochemistry

Da — 若无特别说明,分子量单位默认为道尔顿。   Mw — 重均分子量。   Mn — 数均分子量。

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阿拉丁科学.《Analysis and handling of IHC common problems》. 阿拉丁知识库,更新于 2024年10月30日。 https://www.aladdin-e.com/zh_cn/faqs/analysis-and-handling-of-ihc-common-problems-en.html
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