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. Author manuscript; available in PMC: 2011 May 11.
Published in final edited form as: Biochem Biophys Res Commun. 2002 Apr 26;293(1):463–469. doi: 10.1016/S0006-291X(02)00238-3

Figure 2. Co-immunoprecipitation of p70S6K and MAPK proteins.

Figure 2

(A) Neutrophils (1×107 cells/ml) were incubated in the presence (+) or the absence (−) of 300 pM GM-CSF for 5 minutes. Lysates were immunoprecipitated (I.P.) with anti-MAPK (ERK1+2) antibodies and Western blots (W.B.) derived from immunocomplex beads were subjected to probing with anti-MAPK (left panel) followed by stripping of the antibody and re-probing with anti-PY (middle panel). Shown in both cases are the regions of the blot around the 44 kDa protein marker. Finally, the blot was stripped and re-probed with anti-p70S6K antibodies (right panel, showing the region of the blot around the 71 kDa marker). (B) Neutrophils immunoprecipitated with anti-p70S6K (C-18) antibodies followed by Western blotting with the same antibody (left panel, showing the region of the blot around the 71 kDa protein marker) and subsequent re-probing with anti-MAPK (middle panel, showing the 44-kDa region of the blot). Finally, the blot was probed again with anti-PY (PY20) (right panel, shown again is the region ~ 44 kDa). Results are typical among three other experiments.