I am looking for an answer related to western blot, we have mouse eye tissues which is formalin fixed and then stored in 75% ethanol, Can I use it successfully for western blot?
Hello Nisa Sindhi
The reliability of protein profiles will depend on the effects of formalin fixation. The availability and quality of proteins in a tissue context will depend on several factors like type of fixative, time of fixation, temperature during storage as well as protocols used for protein extraction and detection.
Formalin may not be the optimal fixative. Its deleterious effects on DNA and RNA are well-known. Proteins are altered by the cross-linking mechanism, mainly in the tertiary and quaternary structures, and formalin fixation could render epitopes inaccessible to antibody recognition.
Unfortunately, an optimal fixative that could preserve the biological material for analysis using other technical approaches such as Western Blotting does not exist. However, the use of non-aldehyde-based fixatives could be more suitable for extraction of proteins in obtaining high protein yield for Western blot analysis.
You may want to refer to the paper attached below for more information.
Article The Impact of Tissue Fixatives on Morphology and Antibody-ba...
Best.
Antigens got fixed in formalin will not bind to antibody. not possible to do western.
You can snap freeze in liquid nitrogen and thereafter, store frozen tissues at -80 which can then be pulverized and lysates prepared for WB. Formalin is not recommended.
The integrity of the protein will be compromised due tobthe tissue fixation method. Tissues can either be snapfrozen immediately and stored in -80 (depending on how soon you want to extract your proteins)
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