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Anti-WAVE2, clone ZS33-1, Cat. No. MABT1320, is a mouse monoclonal antibody that detects Wiskott-Aldrich syndrome protein family member 2 and is tested for use in Immunofluorescence, Immunoprecipitation, and Western Blotting.
More>>Anti-WAVE2, clone ZS33-1, Cat. No. MABT1320, is a mouse monoclonal antibody that detects Wiskott-Aldrich syndrome protein family member 2 and is tested for use in Immunofluorescence, Immunoprecipitation, and Western Blotting. Less<<
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Übersicht
Replacement Information
Description
Catalogue Number
MABT1320-25UL
Description
Anti-WAVE2 Antibody, clone ZS33-1
Alternate Names
Wiskott-Aldrich syndrome protein family member 2
WASP family protein member 2
Protein WAVE-2
Verprolin homology domain-containing protein 2
Background Information
Wiskott-Aldrich syndrome protein family member 2 (UniProt: Q9Y6W5; also known as WASP family protein member 2, Protein WAVE-2, Verprolin homology domain-containing protein 2) is encoded by the WASF2 (also known as WAVE2) gene (Gene ID: 10163) in human. WAVE-2 is expressed in all tissues with strongest expression observed in placenta, lung, and peripheral blood leukocytes. Its expression lacks in skeletal muscle. It is required for actin polymerization at the leading edge of migrating cells. It is shown to promotes the formation of actin filaments. WAVE-2 is a component of the WAVE2 complex composed that is made of ABI1, CYFIP1/SRA1, NCKAP1/NAP1 and WAVE-2. This complex regulates actin filament reorganization via its interaction with the Arp2/3 complex. It binds and activates the Arp2/3 complex via the C-terminal domain. It also has a WH2 domain (aa 436-453) that is required for its interaction with actin. Abl Interactor 1 (Abi1) is reported to interact directly with the WH2 domain of WAVE2 that increases its actin polymerization activity and mediates the assembly of WAVE2 complex. WAVE-2 serves as a downstream effector molecule that is involved in the transmission of signals from tyrosine kinase receptors and small GTPases to the actin cytoskeleton. It is shown to regulate integrin clustering leading to high affinity binding of integrins to their ligands. WAVE2-depleted cells display a substantial reduction in adhesion phenotype following T cell activation. It exists in a basal inactive state and is activated by multiple factors. The three requirements for its activation include its interactions with prenylated Rac-GTP, acidic phospholipid, and a specific phosphorylation state. (Ref.: Pauker, MH., et al., (2014). J. Biol. Chem. 289(50); 34503-34519; Innocenti, M., et al., (2004). Nat. Cell Biol. 6(4); 319-327).
References
Product Information
Format
Purified
Presentation
Purified mouse monoclonal antibody IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Applications
Application
Anti-WAVE2, clone ZS33-1, Cat. No. MABT1320, is a mouse monoclonal antibody that detects Wiskott-Aldrich syndrome protein family member 2 and is tested for use in Immunofluorescence, Immunoprecipitation, and Western Blotting.
Key Applications
Immunofluorescence
Immunoprecipitation
Western Blotting
Application Notes
Western Blotting Analysis: A 1:500 dilution from a representative lot detected WAVE2 in HeLa cell lysates.
Western Blotting Analysis: A representative lot detected WAVE2 in Western Blotting applications (Innocenti, M., et. al. (2004). Nat Cell Biol. 6(4):319-27).
Immunofluorescence Analysis: A representative lot detected WAVE2 in Immunofluorescence applications (Marchesin, V., et. al. (2015). PLoS One. 10(3):e0121747).
Immunoprecipitation Analysis: A representative lot detected WAVE2 in Immunoprecipitation applications (Innocenti, M., et. al. (2004). Nat Cell Biol. 6(4):319-27).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Biological Information
Immunogen
GST-tagged recombinant fragment corresponding to 77 amino acids from the C-terminal region of human Wiskott-Aldrich syndrome protein family member 2 (WAVE-2).
Epitope
C-terminus
Clone
ZS33-1
Concentration
0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Host
Mouse
Specificity
Clone ZS33-1 is a mouse monoclonal antibody that detects Wiskott-Aldrich syndrome protein family member 2 (WAVE-2). It targets an epitope within the 77 amino acids from the C-terminal VCA domain.
~70 kDa observed; 54.28 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Western Blotting in lysates from HeLa cells and HeLa cells with WVE2 knockout..
Western Blotting Analysis: A 1:250 dilution of this antibody detected Wiskott-Aldrich syndrome protein family member 2 (WAVE-2) in lysate from HeLa cells, but not in HeLa cells with WAVE2 knockout.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
ARF6 promotes the formation of Rac1 and WAVE-dependent ventral F-actin rosettes in breast cancer cells in response to epidermal growth factor. Marchesin V, Montagnac G, Chavrier P. PLoS One
10(3)
e0121747
2015
Coordination between actin cytoskeleton assembly and localized polarization of intracellular trafficking routes is crucial for cancer cell migration. ARF6 has been implicated in the endocytic recycling of surface receptors and membrane components and in actin cytoskeleton remodeling. Here we show that overexpression of an ARF6 fast-cycling mutant in MDA-MB-231 breast cancer-derived cells to mimick ARF6 hyperactivation observed in invasive breast tumors induced a striking rearrangement of the actin cytoskeleton at the ventral cell surface. This phenotype consisted in the formation of dynamic actin-based podosome rosette-like structures expanding outward as wave positive for F-actin and actin cytoskeleton regulatory components including cortactin, Arp2/3 and SCAR/WAVE complexes and upstream Rac1 regulator. Ventral rosette-like structures were similarly induced in MDA-MB-231 cells in response to epidermal growth factor (EGF) stimulation and to Rac1 hyperactivation. In addition, interference with ARF6 expression attenuated activation and plasma membrane targeting of Rac1 in response to EGF treatment. Our data suggest a role for ARF6 in linking EGF-receptor signaling to Rac1 recruitment and activation at the plasma membrane to promote breast cancer cell directed migration.
Abi1 is essential for the formation and activation of a WAVE2 signalling complex. Innocenti M, Zucconi A, Disanza A, Frittoli E, Areces LB, Steffen A, Stradal TE, Di Fiore PP, Carlier MF, Scita G. Nat Cell Biol
6(4)
319-27
2004
WAVE2 belongs to a family of proteins that mediates actin reorganization by relaying signals from Rac to the Arp2/3 complex, resulting in lamellipodia protrusion. WAVE2 displays Arp2/3-dependent actin nucleation activity in vitro, and does not bind directly to Rac. Instead, it forms macromolecular complexes that have been reported to exert both positive and negative modes of regulation. How these complexes are assembled, localized and activated in vivo remains to be established. Here we use tandem mass spectrometry to identify an Abi1-based complex containing WAVE2, Nap1 (Nck-associated protein) and PIR121. Abi1 interacts directly with the WHD domain of WAVE2, increases WAVE2 actin polymerization activity and mediates the assembly of a WAVE2-Abi1-Nap1-PIR121 complex. The WAVE2-Abi1-Nap1-PIR121 complex is as active as the WAVE2-Abi1 sub-complex in stimulating Arp2/3, and after Rac activation it is re-localized to the leading edge of ruffles in vivo. Consistently, inhibition of Abi1 by RNA interference (RNAi) abrogates Rac-dependent lamellipodia protrusion. Thus, Abi1 orchestrates the proper assembly of the WAVE2 complex and mediates its activation at the leading edge in vivo.
