VCA-GST
Human recombinant WAVE VCA-domain, GST-tagged
Cat. #: 8416-01
Applications
VCA-GST is a constitutively active nucleation-promoting factor (NPF) fragment derived from WAVE/SCAR proteins of the WASP family. It is used to activate the Arp2/3 complex in defined in vitro systems to generate branched actin networks, enabling quantitative polymerization assays (e.g., pyrene-actin fluorimetry) and minimal reconstitutions of Arp2/3-driven actin assembly.
The V (WH2/verprolin) element recruits G-actin, while the CA region engages and activates Arp2/3; together they accelerate branch nucleation from pre-existing filaments under controlled buffer conditions. The GST tag supports affinity handling and immunodetection with anti-GST reagents.
Description
| Protein | VCA-GST |
| Origin | recombinant, human (E. coli expression) |
| Molecular mass | ~37.5 kDa |
| Protein description |
The VCA domain (verprolin/WH2–central–acidic) is the conserved C-terminal output module of WASP-family NPFs, including WAVE/SCAR. In isolation, VCA is active: it binds actin monomers via the V (WH2) region and engages the Arp2/3 complex via the CA region, thereby stimulating Arp2/3-dependent nucleation of branched actin filaments. This product is supplied as a GST-tagged VCA fusion expressed in E. coli and purified by GST affinity chromatography followed by gel filtration. Functional activity is confirmed by activation of the Arp2/3 complex in fluorimetric assays using 10% pyrene actin. |
| Actin interaction | Binds G-actin (V/WH2) and activates the Arp 2/3 protein complex (Cat. #: 8413-01) to drive branched actin nucleation. |
Properties
| Form | Lyophilized, ready-to-use. |
| Quantity per unit | 1 × 250 µg |
| Buffer | 150 mM NaCl, 20 mM HEPES pH 7.4, 1 mM DTT, 5% sucrose (upon reconstitution with 250 µl ultrapure water to obtain a 1.0 mg/ml solution) |
| Purity | >99% by scanning densitometry from Coomassie G-250 stained SDS gels. |
| Purification notes | Expressed in E. coli; purified on GST-Sepharose and further polished by gel filtration. Detectable with anti-GST antibodies in standard immunological detection systems. |
| Protein concentration | Determined at OD280 (0.1% = 1.25). |
| Storage instructions | Store at -70°C upon arrival. Keep dissolved protein on ice during use; for longer storage of reconstituted protein, store at -20°C in glycerol or CryoProtec (Cat. #: 6011-01). Avoid repeated freeze/thaw cycles. |
| Shipping conditions | At ambient temperature. Upon delivery store at -70°C. |
| Remarks | For research use only. Not for use in human or veterinary diagnostic or therapeutic applications. |
| CAS no. |
Proteins from Hypermol® are made of the ultrapure reagents in Milli-Q™ water, as described in our publications.
Further Information
Material and Safety Data Sheet
Protein Structures on RCSB (WAVE)
References
Reconstitution of contractile actomyosin rings in vesicles.
Litschel T, Kelley CF, Holz D, Adeli Koudehi M, Vogel SK, Burbaum L, Mizuno N, Vavylonis D, Schwille P. Nat Commun. 2021 Apr 15;12(1):2254. doi: 10.1038/s41467-021-22422-7.
Initiation of DNA replication requires actin dynamics and formin activity.
Parisis N, Krasinska L, Harker B, Urbach S, Rossignol M, Camasses A, Dewar J, Morin N, Fisher D. EMBO J. 2017 Nov 2;36(21):3212-3231. doi: 10.15252/embj.201796585.
Structure and control of the actin regulatory WAVE complex.
Chen Z, Borek D, Padrick SB, Gomez TS, Metlagel Z, Ismail AM, Umetani J, Billadeau DD, Otwinowski Z, Rosen MK. Nature. 2010 Nov 25;468(7323):533-8. doi: 10.1038/nature09623.
Product DataSheetMaterial and Safety Data SheetProtein Structures on RCSB (WAVE)
Reconstitution of contractile actomyosin rings in vesicles.
Litschel T, Kelley CF, Holz D, Adeli Koudehi M, Vogel SK, Burbaum L, Mizuno N, Vavylonis D, Schwille P.Nat Commun. 2021 Apr 15;12(1):2254. doi: 10.1038/s41467-021-22422-7.
Initiation of DNA replication requires actin dynamics and formin activity.
Parisis N, Krasinska L, Harker B, Urbach S, Rossignol M, Camasses A, Dewar J, Morin N, Fisher D.
EMBO J. 2017 Nov 2;36(21):3212-3231. doi: 10.15252/embj.201796585. Epub 2017 Oct 5.
Structure and control of the actin regulatory WAVE complex.
Chen Z, Borek D, Padrick SB, Gomez TS, Metlagel Z, Ismail AM, Umetani J, Billadeau DD, Otwinowski Z, Rosen MK.
Nature. 2010 Nov 25;468(7323):533-8. doi: 10.1038/nature09623.
