Western blot analysis of extracts from HeLa, Jurkat and COS cells using ARP3 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 BSA, 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.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
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: 10
ARP3 Antibody detects endogenous levels of total ARP3 protein. The antibody does not cross-react with endogenous levels of total ARP2 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of human ARP3. Antibodies are purified using protein A and peptide affinity chromatography.
Actin nucleation, the formation of new actin filaments from existing filaments, affects actin filament structure during cell motility, division, and intracellular trafficking. An important actin nucleation protein complex is the highly conserved ARP2/3 complex, consisting of ARP2, ARP3, and ARPC1-5. The ARP2/3 complex promotes branching of an existing actin filament and formation of a daughter filament following activation by nucleation-promoting factors, such as WASP/WAVE or cortactin (1). The formation of podosomes, small cellular projections that degrade the extracellular matrix, is enhanced by ARP2/3 complex action. ARP2/3 competes with caldesmon, an actin binding protein shown to negatively affect podosome formation (2). Along with N-WASP, the ARP2/3 complex regulates nuclear actin filament nucleation and controls actin polymerization during transcription (3).
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