Actin is an abundant cytoskeletal protein found in all cells (True, 1990). The protein's 42 kD peptide chain assumes two physical forms: globular actin, which may serve as a cytoplasmic storage pool, and fibrous actin, which, in conjunction with myosin, generates muscle contraction (True, 1990). In non-muscle cells, actin appears to be involved in a variety of functions, such as cell motility, exocytosis, and phagocytosis (True, 1990). Distribution of the six known isoforms of actin - four muscle actins (alpha-skeletal, alpha-vascular smooth, alpha-cardiac, and gamma-enteric smooth) and two cytoplasmic actins (alpha and gamma) is tissue specific (Otey, 1986; Lessard, 1988).
Applications
Suitable for use in Immunofluorescence, Immunocytochemistry, Immunohistochemistry, Western Blot and ELISA. Other applications have not been tested.
Recommended Dilutions
Immunofluorescence: 1:500 using NIH/3T3 cells. Positive staining pattern reflects both membrane and cytolasmic staining. Immunocytochemistry: 10ug/ml using methanol fixed mouse 3T3 cells Immunohistochemistry (Paraffin): 10ug/ml using paraffin-embedded, 4% formaldehyde, 3% glutaraldehyde, sodium cacodylate treated sections Western Blot: 1:5000 using HEK293 lysates ELISA: strongly reactive with the cytoplasmic actin and shows a significant binding to gizzard, skeletal, arterial and cardiac actins. Also shows a significant binding to both Dictyostelium discoidum and Physarum polycephalum. Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Purified chicken gizzard actin. Epitope: Appears to be specific for the N-terminal two-thirds of the actin molecule, possibly near aa50-70.
Form
Supplied as a liquid. No preservative added.
Specificity
Recognizes all six isoforms of vertebrate actin, globular (G) and fillimentous (F) forms. This clone is a pan-actin antibody that binds to an epitope in a highly conserved region of actin. Labels myotubes and stains myoblasts and fibroblasts. Does not interfere with actin polymerization to form filaments, at a ratio as high as one antibody per two actin monomers. Species Crossreactivity: All vertebrates as well as Dictyostelium discoideum and Physarum polycephalum.
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