| Description |
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. |