Western blot analysis of extracts from various cell lines using LASP1 Antibody.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 263
LASP1 Antibody detects endogeneous levels of total LASP1 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala191 of human LASP1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
LASP1 is a cytoskeletal scaffold protein belonging to the LIM protein subfamily (1,2). LASP1 consists of an N-terminal LIM domain, followed by two nebulin repeats, and a C-terminal SH3 domain (1,3). The nebulin repeats interact with actin, while the SH3 domain interacts with palladin (4,5), suggesting LASP1 functions as an actin-binding protein, possibly in cytoskeletal organization. LASP1 has been shown to localize to focal adhesions, lamellipodia, and membrane ruffles (6-8) and might be involved in membrane migration. Overexpression of LASP1 has been associated with metastatic cancers, such as breast and ovarian cancer (2). In these cases, membrane, cytoplasmic, and nuclear localization of LASP1 in the tumor cell has been reported, suggesting LASP1 involvement in membrane and nuclear signaling (9,10).
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