Actin, a ubiquitous eukaryotic protein, is the major component of the cytoskeleton. At least six isoforms are known in mammals. Nonmuscle b- and g-actin, also known as cytoplasmic actin, are predominantly expressed in nonmuscle cells, controlling cell structure and motility (1). a-cardiac anda-skeletal actin are expressed in striated cardiac and skeletal muscles, respectively; two smooth muscle actins, a- and g-actin, are found primarily in vascular smooth muscle and enteric smooth muscle, respectively. These actin isoforms regulate the contractile potential of muscle cells (1). Actin exists mainly as a fibrous polymer, F-actin. In response to cytoskeletal reorganizing signals during processes such as cytokinesis, endocytosis, or stress, cofilin promotes fragmentation and depolymerization of F-actin, resulting in an increase in the monomeric globular form, G-actin (2). The Arp2/3 complex stabilizes F-actin fragments and promotes formation of new actin filaments (2). It has been reported that actin is hyperphosphorylated in primary breast tumors (3). Cleavage of actin under apoptotic conditions has been observed in vitro and in cardiac and skeletal muscle (4-6). Actin cleavage by caspase-3 may accelerate ubiquitin/proteosome dependent muscle proteolysis (6).
Applications:
Suitable for use in Immunofluorescence, Flow Cytometry, Western Blot, Immunohistochemistry and Immunocytochemistry. Other applications not tested.
Recommended Dilution:
Immunofluorescence (IF-IC): 1:100. Methanol permeabilization is required.
Flow Cytometry: 1:100
Western Blot: 1:1000
Immunohistochemistry (Paraffin): 1:200. Antigen retrieval with citrate buffer is recommended.
Immunohistochemistry (Frozen): 1:200. Fixation in 10% Neutral buffered formalin is recommended.
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.