Description_x000D_ General description_x000D_ Actin is a highly conserved protein that is a major component of both the cytoskeletal and contractile structures in all cell types. It varies in amount, being related to the type of differentiation and to the functional state of cells and tissues. Actin can be found in two different forms of aggregation, the globular or the fibrillar state, and at least six distinct isoforms occur in vertebrates. The actins exhibit over 90% sequence homology, but each isoform has a unique NH2-terminal sequence. The isoforms are comprised of three α actins (skeletal, cardiac, smooth), one β actin (β-non-muscle) and two γ actins (γ smooth muscle and γ non-muscle). The human γ-actin gene is mapped to human chromosome 17._x000D_ The antibody recognizes an epitope located on the C-terminal end of actin conserved in all actin isoforms. It specifically labels actin in a wide variety of tissues and species. The epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding. Zinc-formalin, B5, ethanol, methacarn, Brunnel′s or Bouin′s solutions may also be used as fixatives._x000D_ Monoclonal Anti-Actin (mouse IgG2a isotype) is derived from the AC-40 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from an immunized mouse. A synthetic actin C-terminal peptide, attached to Multiple Antigen Peptide (MAP) backbone was used as the immunogen. The isotype is determined using Mouse Monoclonal Antibody Isotyping Reagents, Catalog No. ISO2_x000D_ The antibody recognizes an epitope located on the C-terminal end of actin conserved in all actin isoforms. It specifically labels actin in a wide variety of tissues and species. The epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding. Zinc-formalin, B5, ethanol, methacarn, Brunnel′s or Bouin′s solutions may also be used as fixatives._x000D_ Specificity_x000D_ Cross-reaction of monoclonal Anti-Actin antibody has been observed with human, bovine, sheep, goat, pig, rabbit, dog, mouse, rat, guinea pig, hamster, chicken, carp, viper, frog and snail. The antibody may be used in staining of methanol-fixed, frozen sections._x000D_ Immunogen_x000D_ synthetic actin C-terminal peptide, attached to a Multiple Antigen Peptide (MAP) backbone._x000D_ Application_x000D_ Monoclonal anti-actin antibody was used for western blot analysis of Cos-7 cell lysates to ensure equal protein loading._x000D_ Mouse monoclonal anti-actin antibody can be used for western blot assays. Furthermore, the product can also be used for immunocytochemistry (10-20μg/mL, using chicken or human fibroblasts by methanol/acetone fixation), microarray, immunohistochemistry and indirect ELISA._x000D_ Monoclonal Anti-Actin antibody produced in mouse has been used in the detection of actin in human bronchial epithelial cells using immunohistochemical assays. It has also been used in immunofluorescence detection in cardiomyocytes._x000D_ Western blot analysis of MDCK cell lysates were performed using monoclonal anti-actin antibody as a primary antibody._x000D_ Physical form_x000D_ Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide._x000D_ Storage and Stability_x000D_ For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. Storage in “frost-free” freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours._x000D_ Disclaimer_x000D_ Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals._x000D_ Packaging_x000D_ 200 μL in glass insert_x000D_ Biochem/physiol Actions_x000D_ Actin and myosin are constituents of many cell types and are involved in cellular process including locomotion, secretion, cytoplasmic streaming, phagocytosis, and cytokinesis. The N-terminal region of actin appears to be a major antigenic region, and may be involved in the interaction of actin with other proteins such as myosin. Among the six isoforms of actin, four of them represent the differentiation markers of muscle tissues and two are found practically in all cells. There are three α-actins (skeletal, cardiac, and smooth muscle), one β-actin (β-nonmuscle), and two γ-actins (γ-smooth muscle and γ-nonmuscle). Actin isoforms show >90% overall sequence homology, but only 50–60% homology in their NH2-terminal residues._x000D_ The actin in cells of various species and tissues are very similar in their immunological and physical properties characterized by electrophoresis and amino acid sequence analysis. They mediate cytoskeletal remodeling during T cell activation. Mutation in the γ-actin and β-actin gene is associated with a developmental disorder called Baraitser-Winter syndrome. Mutation in the γ-actin leads to hearing loss._x000D_ Other Notes_x000D_ To view an Actin antibody selection guide, please visit www.sigmaaldrich.com/actin.