Fig. 2.

Characterization of γ-actin-specific antibodies. (A) Gels equally loaded with purified sarcomeric α-actin, platelet actin (a 4:1 mixture of β- and γ-isoforms), and brain γ-actin were either stained with Coomassie blue (CB) or transferred to nitrocellulose and Western-blotted with mAbs specific for α- or β-actin. (B) Western blots were stained with affinity-purified pAbs raised against the unique amino-terminal peptide sequence of cytoplasmic γ-actin or mAbs from mice immunized with both purified brain γ-actin and the amino-terminal sequence of cytoplasmic γ-actin. Where indicated, 10 μg/ml of either the amino-terminal γ-actin or β-actin peptide was preincubated with the γ-actin antibodies. (C) Purified platelet β/γ-actin was resolved by 2D electrophoresis and transferred to poly(vinylidene difluoride), and the blot was serially probed with γ-actin pAbs, γ-actin mAbs, and finally β-actin mAbs. (D) Ten-micrometer transverse and longitudinal cryosections of control tibialis anterior muscle stained with the γ-actin mAb. (Scale bar: 50 μm.)