The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500 - 1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).
Use at an assay dependent concentration.
Controls reversibly actin polymerization and depolymerization in a pH-sensitive manner. It has the ability to bind G- and F-actin in a 1:1 ratio of cofilin to actin. It is the major component of intranuclear and cytoplasmic actin rods.
Widely distributed in various tissues.
Belongs to the actin-binding proteins ADF family. Contains 1 ADF-H domain.
Phosphorylated on Ser-3 in resting cells.
Nucleus matrix. Cytoplasm > cytoskeleton. Almost completely in nucleus in cells exposed to heat shock or 10% dimethyl sulfoxide.
Western blot - Anti-Cofilin (phospho S3) antibody (ab47281)
All lanes : Anti-Cofilin (phospho S3) antibody (ab47281) at 1/500 dilution
Lane 1 : Extracts from COLO205 cells serum starved Lane 2 : Extracts from COLO205 cells
Lysates/proteins at 30 µg per lane.
Predicted band size : 18 kDa Observed band size : 18 kDa Preparation of positive control: when the cells were at the log phase and 80-90% confluent, the culture media was drained. Cells were washed with fresh basic media (no serum) once. Add fresh basic media to the vessels and starve the cells for 18-24 hours. Drain the cultured medium, then add 20% serum to the vessels for 15 mins.