Hey,
I am thinking of extracting overall protein from formaldehyde-fixed cells via the "boiling" method, as described by Sadick et al. 2016, 2017 (see below), and further process the samples in the Pierce BCA assay. Does anyone have practical experiences with this? Are there any pitfalls? Especially, I am wondering about the durability of multi-well plates at a temperature of 100C (polystyrene melting point approx 240C).
Best
Sebastian
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There are reactions of formaldehyde (CH2O) with the terminal amino groups (-NH2) and the polar functions of the side chains of amino acids.
Lysine is an amino acid with a primary amino group at the end, along four methylene residues - (CH2) -, making this function particularly accessible to formalin molecules.
The chemical reaction between the polar group of amino acids (with a labile proton) takes place by addition to formaldehyde. This chemical reaction is complex.
Fixation with formaldehyde will affect your BCA quantification and you will end up getting skewed values that will affect your SDS-PAGE outcomes.
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