The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 19 kDa (predicted molecular weight: 19 kDa).
Use a concentration of 1 µg/ml.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
FunctionControls 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.
Tissue specificityWidely distributed in various tissues.
Sequence similaritiesBelongs to the actin-binding proteins ADF family. Contains 1 ADF-H domain.
Post-translational modificationsPhosphorylated on Ser-3 in resting cells.
Cellular localizationNucleus matrix. Cytoplasm > cytoskeleton. Almost completely in nucleus in cells exposed to heat shock or 10% dimethyl sulfoxide.
IHC image of Cofilin staining in human liver carcinoma FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab42475, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Western blot - Cofilin antibody (ab42475)
All lanes : Anti-Cofilin antibody - Loading Control (ab42475) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 19 kDa Observed band size : 19 kDa Additional bands at : 68 kDa. We are unsure as to the identity of these extra bands.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Cofilin antibody - Loading Control (ab42475)This image is courtesy of an abreview submitted by Sophie Pezet, ESPCI, France
IHC-FoFr image of Cofilin (ab42475) staining on Rat hippocampus sections. The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.