Actin, pan

Actin, an ubiquitous protein in eukaryotes, is the major component of the cytoskeleton. At least six isoforms are known in mammals. Nonmuscle beta- and gamma-actin, also known as cytoplasmic actin, are predominantly expressed in nonmuscle cells, controlling cell structure and motility. Alpha-cardiac and alpha-skeletal actin are expressed in striated cardiac and skeletal muscles, respectively; two smooth muscle actins, alpha- and gamma-actin, are found primarily in vascular smooth muscle and enteric smooth muscle, respectively. These actin isoforms regulate contractile potentials for muscle cells. 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. The Arp2/3 complex stabilizes F-actin fragments and promotes formation of new actin filaments. It has been reported that actin is hyperphosphorylated in primary breast tumors. Cleavage of actin under apoptotic conditions has been observed in vitro and in cardiac and skeletal muscle. Actin cleavage by caspase-3 may accelerate ubiquitin/proteosome dependent muscle proteolysis.\n\nApplications:\nSuitable for use in Immunoprecipitation, Western Blot and Immunohistochemistry. Other applications have not been tested.\n\nRecommended Dilution: \nImmunoprecipitation (Denatured verified) (Use Protein G): 2ug/mg protein lysate\nWestern Blot: 0.5ug/ml for 2hrs at RT\nImmunohistochemistry (formalin/paraffin): 1:200 for 30 min \nRequires boiling tissues sections in 10mM citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20min.\nOptimal dilutions to be determined by the researcher.\nPositive Control: \nHeLa cells and skeletal muscle. \n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

A0760-51

Size

250ul

Applications

IHC, IP, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat, Bov, Can, Ch/Bird, Prc, Rabbit

Form

Supplied as a liquid in 10mM PBS, pH 7.4, 0.2% BSA, 15mM sodium azide before the addition of glycerol to 40%. Also available without BSA and azide. See A0760-51X

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG1,k

References

1. Lessard J. (1988) Two Monoclonal antibodies to actin: One muscle reactive and one generally reactive. Cell Motility and Cytoskeleton 10: 349- 362.

Additional Info

Recognizes all six known vertebrate isoforms of actin (MW~42kD) and two cytoplasmic actins (beta, gamma) which are highly homologous to one another but which differ from the muscle actins at about 25aa. A0760-51 is highly recommended for monitoring total protein load on Western blots. Species Crossreactivity: Human, bovine, mouse, canine, porcine, rabbit, rat, chicken, Dictyostelium discoideum, Physarum polycephalum.