Hi all, Will it affect antibody staining if paraffin tissue is fixed for 3-4 days?
Yes, extended fixation of paraffin-embedded tissue for 3–4 days, particularly in formalin-based fixatives, can affect antibody staining. Over-fixation leads to excessive crosslinking of proteins, which may mask epitopes and reduce antibody binding efficiency, resulting in weaker or inconsistent immunostaining.
However, this effect can often be mitigated by optimizing the antigen retrieval method (e.g., using citrate buffer pH 6.0 or EDTA pH 9.0 with heat-induced retrieval). It is advisable to validate retrieval conditions for each antibody used. If possible, standardize fixation time (e.g., 24–48 hours) to ensure consistent staining outcomes across samples.
Yan Yan Zhao ,
If paraffin tissue is fixed for 3-4 days, it will affect antibody staining.
Usually, tissue fixation time is 1-2 days, and the specific length of time needs to be determined according to the size of the tissue block.
Too long a fixation time may cause excessive cross-linking of proteins, which may lead to staining failure or high staining background.
The answer to this question comes from MedChemExpress (MCE) Technical Support.
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