Contribution of cell culture additives to the two-dimensional protein patterns of mouse macrophages.
Low levels of fetal calf serum (FCS), used as protein supplement in cell culture medium, were traced in preparations of primary murine macrophages (bone-marrow-derived macrophages (BMM) and peritoneal macrophages (PM)). Main components of this common additive were mapped in 2-DE by means of differential image gel electrophoresis and immunoblotting. Additional washing steps in cell preparation helped to decrease the levels of the four highest abundance foetal serum proteins (serum albumin (SA), alpha1-fetoprotein (AFP), alpha1-antitrypsin (alpha1AT) and transferrin (Tf)) to less than 1% of total protein. Macrophage spot pattern was recorded in parallel and showed little variation. Results presented are supposed to be of general interest for cell preparations with similar background.
Miller
Ingrid
I
Institute of Medical Chemistry, University of Veterinary Medicine, Vienna, Austria. ingrid.miller@vu-wien.ac.at
Radwan
Marta
M
Strobl
Birgit
B
Müller
Mathias
M
Gemeiner
Manfred
M
D000818:Animals; D016477:Artifacts; D002417:Cattle; D018929:Cell Culture Techniques; D003470:Culture Media; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005312:Fetal Blood; D008264:Macrophages / Q000737:chemistry*; D017737:Macrophages, Peritoneal / Q000737:chemistry*; D051379:Mice; D020543:Proteome / Q000737:chemistry*
D003470:Culture Media; D020543:Proteome