Actin

Four muscle actins have been identified: skeletal (alpha), cardiac (alpha), vascular smooth muscle (alpha) and enteric smooth muscle (gamma) actin. Also reacts with both G & F 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.\n\nApplications:\nSuitable for use in Indirect Immunofluorescence, Western Blot, Iodination and Solid Phase Binding. Other applications have not been tested.\n\nRecommended Dilution:\nIndirect immunofluorescence: 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 (6um)-quick frozen in isopentane, slides treated with gelatin and formaldehyde. \n\nImmunoblots: 1:100-1:1000; On muscle homogenates subject to SDS-PAGE, reacts relatively uniformly with a 43kD 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.\n\nSolid Phase Binding Assay: 1:800-1:1000; In an ELISA, is 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. \nOptimal dilutions to be determined by the researcher.\n\nAffinity Constant: \n3-15 x 10e8 L/mole (gizzard > cardiac = skeletal muscle actin)\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol (40-50%), 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

A0759-06

Size

100ul

Applications

ELISA, IF, WB

Hosts

Mouse

Reactivities

Ch/Bird

Form

Supplied as a liquid with 0.01% sodium azide.

P Type

Mab

Purity

Ascites

Isotype

IgG1,k

References

1. Herman, I.M. and T.D. Pollard, J. Cell. Biol. 80: 509-520 (1979). 2. Otey, C.A., et al., J. Cellular Biochem. 34: 113-124 (1987). 3. Lessard, et al., Analyt. Biochem. 94: 140-149 (1979). 4. Cell Motility and the Cytoskeleton 10: 349-362 (1988). 5. Beer-Stoltz, D. and G.K. Michalopoulous, J. Cellular Biochem. 55: 445-464 (1994). 6. Wine, R. and R. Chapin, J. Androl. 20: 198-213 (1999). 7. Mitsui, K., et al., J. Neurosci. 22: 9,267-9,277 (2002). 8. Shou, J., et al., Oncogene 21: 878-889 (2002).

Additional Info

Appears to be specific for the N-terminal two-thirds of the actin molecule (possibly near residues 50-70 in primary sequence). Reacts with every actin form tested. Species Crossreactivity: Reacts with all animal species and cell types with an actin form, including plant actin.