The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. The detection limit for recombinant tagged WASF2 is approximately 0.1ng/ml as a capture antibody.
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 54 kDa).
Use 0.1-1µg for 106 cells. ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
FunctionDownstream effector molecule involved in the transmission of signals from tyrosine kinase receptors and small GTPases to the actin cytoskeleton. Promotes formation of actin filaments. Part of the WAVE complex that regulates lamellipodia formation. The WAVE complex regulates actin filament reorganization via its interaction with the Arp2/3 complex.
Tissue specificityExpressed in all tissues with strongest expression in placenta, lung, and peripheral blood leukocytes, but not in skeletal muscle.
Sequence similaritiesBelongs to the SCAR/WAVE family. Contains 1 WH2 domain.
DomainBinds and activates the Arp2/3 complex via the C-terminal domain. Interacts with actin via the WH2 domain.
Cellular localizationCytoplasm > cytoskeleton. Cell projection > lamellipodium. At the interface between the lamellipodial actin meshwork and the membrane.
Putative Wiskott Aldrich syndrome protein family member 4 antibody
SCAR 2 antibody
Suppressor of cyclic-AMP receptor (WASP family) antibody
Verprolin homology domain-containing protein 2 antibody
WASP family protein member 2 antibody
WASP family protein member 4 antibody
WASP family Verprolin homologous protein 2 antibody
WASP-family protein member 2 antibody
WAVE 2 antibody
Wiskott-Aldrich syndrome protein family member 2 antibody
Wiskott-Aldrich syndrome protein family verprolin-homologous protein antibody
Anti-WASF2 antibody images
Western blot - Anti-WASF2 antibody (ab60722)
Predicted band size : 54 kDa
Lane 1: Wild-type HAP1 cell lysate (40 µg) Lane 2: WASF2 knockout HAP1 cell lysate (40 µg) Lane 3: HeLa cell lysate (20 µg) Lane 4: Jurkat cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab60722 observed at 70 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab60722 was shown to recognize WASF2 when WASF2 knockout samples were used, along with additional cross-reactive bands. Wild-type and WASF2 knockout samples were subjected to SDS-PAGE. Ab60722 and ab181602 (loading control to GAPDH) were diluted at 1 µg/ml and 1:10,000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-WASF2 antibody (ab60722)
Overlay histogram showing K562 cells stained with ab60722 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab60722, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in K562 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Western blot - WASF2 antibody (ab60722)
Anti-WASF2 antibody (ab60722) at 1 µg/ml + HeLa cell lysate at 25 µg