General description

Actins are ubiquitous eukaryotic proteins that serve as a multi-functional, basic building blocks of cytoskeletal microfilaments. They play critical roles in a wide range of cellular processes, including cell division, cell migration, chromatin remodeling, trascriptional regulation and vesicle trafficking. These funstions are attributed to their ability to form filaments, which can quickly assemble and disassemble depending upon the needs of the cell. At least six different actin types have been reported in mammals. Although actins show about 90% overall sequence homology, isoforms do not show spatial, temporal and tissue-specific expression patterns and only 50-60% homology is found in their 18 N-terminal residues. Beta and gamma-actins, also known as cytoplasmic actins, are highly conserved in higher animals and are predominantly expressed in non-muscle cells where they control cell structure. Exocytosis, and motility. They are nearly identical proteins and differ only in four amino acids at the N-terminal region. The other four actin isoforms are typically found in specific adult muscle tissue types. Alpha-cardiac and alpha-skeletal actins are expressed in striated cardiac and skeletal muscles, respectively. Alpha and gamma actins are primarily found in vascular smooth muscle and enteric smooth muscles, respectively. It has been shown that under calcium-bound conditions, beta-actin exhibits more dynamic behavior than gamma-actin with higher rates of polymerization and depolymerization. Also, beta- and gamma-actins can readily copolymerize, and the resulting filaments exhibit polymerization and depolymerization rates that vary depending on the ration of beta- to gamma-actin (Lessard, JL.,et al.(1988). Cell Motility Cytoskeleton 10(3); 349-362.

Specificity

MAB1501 is a pan-actin antibody that binds to an epitope in a highly conserved region of actin; therefore, this antibody reacts with all six isoforms of vertabrate actin (Lessard, 1988). Reacts with both globular (G) and fillimentous (F) forms of actin and does not interfere with actin polymerization to form filaments, at a ratio as high as one antibody per two actin monomers. However, this antibody does increase the extent of polymerization when used at a lower ratio of antibody to actin. In addition to labeling myotubes, anti-actin stains myoblasts and fibroblasts. Although clone C4 is prepared as an antibody to chicken gizzard muscles actin, it reacts with actins from all vertebrates, as well as with Dictyostelium discoideum and Physarum polycephalum actins (Lessard, 1988).

To date, all animal species and cell types with an actin form react by indirect immunofluorescence or immunoblot, including plant actin.

Immunogen

Epitope is conserved in all known actins.

Purified chicken gizzard actin (Lessard, 1988).

Application

Indirect immunofluorescence at 1:100:
Tissue culture cells -- fix with formaldehyde, treat with methanol or acetone.
Glycerinated myofibrils -- fix fibers with formaldehyde, treat with cold methanol. Stains I-bands intensely and stress fibers in human fibroblasts.
Cryostat sections (6 µm) -- quick frozen in isopentane, slides treated with gelatin and formaldehyde.

Immunoblots:
1:100-1:1,000 (Otey, 1987):On muscle homogenates subject to SDS-PAGE, reacts relatively uniformly with a 43 kD protein present in skeletal, cardiac, gizzard and aorta tissues. Appears to react with all isoforms of actin found in these preparations and shows a strong reaction with the alpha-actin found in skeletal, cardiac, and arterial muscle.

Iodination (Lessard, 1979).

Solid phase binding assay ELISA:
1:800-1:1,000 dilution from a previous lot was shown to be strongly reactive with 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.

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 working dilutions must be determined by end user.

Reliably and specifically detect actin using this Anti-Actin Antibody, clone C4. This highly published monoclonal antibody is validated for use in ELISA, IC, IF, IH, IH(P) & WB. This mAb is also available as a fluorescent conjugate.

Research Category
Cell Structure

Research Sub Category
Cytoskeletal Signaling

Quality

Routinely evaluated by Western Blot on A431 lysates.

Western Blot Analysis:
1:500 dilution of this lot detected ACTIN on 10 ug of A431 lysates.

Target description

43 kDa

Linkage

Replaces: 04-1040

Physical form

Mouse monoclonal Ascites fluid, with 0.01% sodium azide.

Unpurified

Storage and Stability

Stable at -20°C in undiluted aliquots for up to 12 months from date of receipt. Do not store in a diluted format. Avoid repeated freeze/thaw cycles.

Analysis Note

Control
HeLa whole cell lysate.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

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.

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