Figure 3. Characterization of antibodies that recognize FLNc.

Bacterially expressed FLNc (residues 1915–2446) was left unphosphorylated (no kinase; NK) or maximally phosphorylated with PKBα. Aliquots (50 ng) were spotted on to a nitrocellulose membrane (Schleicher and Schuell, London, U.K.) and immunoblotted using the anti-pSer²²¹³ antibody (α-pS2213) or an antibody that recognizes phosphorylated and unphosphorylated FLNc equally well (α-FLNc) in the presence or absence (‘none’) of the competitor peptides shown. The sequences of these peptides are given in the Materials and methods section. The prefix p denotes a phosphorylated peptide.