Actin is a 43kD highly conserved structural protein. There are three main actin isotypes (alpha,beta and gamma) which show >90% amino-acid (aa) homology between isotypes and >98% homology within members of a particular isotypic group. The majority of the isotype heterogeneity is located in the amino-terminal 30aa. The different actin isotypes have been shown to behave very differently in vitro and in vivo. Recent studies describe differential subcellular localization of gamma-actin and isotype specific binding of actin associated proteins. There are at least six actin isoforms in vertebrates: cardiac alpha-actin and skeletal-alpha actin (expressed in striated muscle), vascular alpha-actin and enteric gamma-actin (expressed in smooth muscle), and cytoplasmic beta- and gamma-actins.
Applications
Suitable for use in Western Blot, Immunohistochemistry and Immunocytochemistry. Other applications not tested.
Recommended Dilutions
Western Blot: 1:500-1:2000. Non-specific bands can occur when A0760-45 is overloaded. Blocking with purified milk proteins preferred over commerical concentrated milk solutions. Detects a 42-43kD band Immunocytochemistry: 1:500-1:2000. Cultured cells fixed with PFA, then permeabilized with methanol. Immunohistochemistry: 1:500-1:2000. Tissues frozen and then fixed with PFA followed by Triton permeabilisation. 10% buffered formalin fixed tissues have also been used 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
Synthetic peptide corresponding EEEIAALVIDNGSGGSG at the N-terminus of gamma actin, conjugated to KLH. Cellular Localization: Cytoplasm and cytoskeleton
Form
Supplied as a liquid.
Specificity
Recognizes human gamma actin. Species Crossreactivity: rat and mouse. Actins are highly conserved proteins that are involved in various types of cell motility, and maintenance of the cytoskeleton.