Mature αS1-casein binds to post-ER membranes of the secretory pathway. A, B. Aliquots of PNS were diluted and incubated in conservative (C) buffer in the absence of saponin (-) or in conservative or non-conservative (NC) buffers in the presence of saponin (+) and centrifuged. Supernatants (S) and pellets (P) were analyzed by SDS-PAGE. A. Coomassie Blue staining. B. Immunoblotting with an antibody against mouse milk proteins. The quantity of mature αS1-casein in the supernatant and the pellet was quantified by densitometry and expressed as percent of the total quantity of the casein (supernatant + pellet). The mean ± s.d. of three independent experiments is shown. C. Aliquots of PNS were subjected to carbonate extraction at pH 11 with saponin in the absence (-) or in the presence (+) of DTT and centrifuged. Supernatants and pellets were analyzed by SDS-PAGE followed by immunoblotting as above. Data are representative of at least three independent experiments. Relative molecular masses (kDa) are indicated. m. αS1-cas: mature αS1-casein.
Le Parc et al. BMC Cell Biology 2010 11:65 doi:10.1186/1471-2121-11-65