Hello,
For many phosphorylation studies of MAP kinases, cells are serum-starved overnight prior to stimulation. Serum-free media such as DMEM 1X is supplemented with 0.5% bovine serum albumin. Is there any drawback to including NEAA in the serum-free media? Is NEAA known to cause phosphorylation of MAP kinases?
Thanks!
Hello David Schad
Addition of NEAA in serum-free media may affect your experimental results. Amino acids are the basic building blocks of proteins and constitute all proteinaceous material of the cell including the protein component of enzymes, receptors, and signaling molecules. Also, some non-essential amino acids can regulate gene expression and cell signaling pathways. Therefore, under normal conditions, NEAA is used as a supplement for cell culture medium to increase cell growth and viability.
Moreover, MAP kinases regulate diverse cellular programs by relaying extracellular signals to intracellular responses, and coordinately regulate cell proliferation, differentiation, motility, and survival. So, I feel you should limit to serum-free media supplemented with 0.5% bovine serum albumin so that the cells do not proliferate but manage to survive overnight prior to stimulation.
Best.
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