Comparison

VCA domain of WASP (GST fusion)

Item no. VCG03-A
Manufacturer Cytoskeleton
Amount 1 x 500 ug
Category
Type Proteins
Specific against other
Purity Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. Samples are >95% pure (Figure 1).
Citations van der Gucht, J., Paluch, E., Plastino, J. and Sykes, C. (2005). Stress release drives symmetry breaking for actin-based movement. Proc. Natl. Acad. Sci. U. S. A. 102, 7847-7852.
ECLASS 10.1 32160409
ECLASS 11.0 32160409
UNSPSC 12352202
Alias Actin, Actin Protein, VCA, WASP, WASP protein, WASP-VCA domain, WASP VCA, WASP VCA domain
Similar products Actin, WASP, VCA, Actin Protein, WASP protein, WASP-VCA domain, WASP VCA, WASP VCA domain
Shipping condition Room temperature
Available
Shipping Temperature
AT
Storage Conditions
On Arrival: 4°C
Delivery Time
1-2 Weeks
FAQs

Question 1: By itself, what is the effect of VCA-domain WASP protein on actin polymerization?


Answer 1: By itself, the VCA-domain WASP protein ((Cat. #VCG03) partial inhibits actin polymerization. When combined with Arp2/3 (RP01), actin polymerization is greatly enhanced. See the associated datasheets for details.


Question 2: What is the optimal concentration of Arp2/3 and VCA-domain WASP protein to modify actin polymerization?


Answer 2: We recommend thatpyrene actin is present at a final concentration of 0.8 &mu, M, Arp2/3 complex at 10 nM (RP01) and the VCA domain of WASP at 400 nM ((Cat. #VCG03). The critical factor here is the actin concentration which must be 0.80 +/-0.2 µ, M in the final reaction. This concentration is just below the critical concentration for assembly and hence will not polymerize on it&rsquo, s own.

Weight (grams)
8
Product Uses
Stimulating Arp2/3 complex activity
Isolating Arp2/3 complex from cell or tissue extract
Material
The VCA (Verprolin, cofilin, acidic) domain of human WASP protein has been expressed in a bacterial expression system as a GST-tagged fusion protein. The protein has been purified by glutathione affinity chromatography and is supplied as a lyophilized powder.When reconstituted with distilled water, the complex is in the following buffer: 20 mM Tris pH 7.5, 25 mM KCl, 1 mM MgCl2 0.5 mM EDTA, 0.2% dextran, 2% sucrose. The molecular weight of the GST-VCA domain protein is approximately43 kDa.
Biological Activity
VCG03 was tested in an actin polymerization assay (Cat. # BK003). In conjunction with Arp2/3 (Cat. # RP01>) it was shown to stimulate actin polymerization 20-fold compared to the control without VCG03. This indicates VCA domain specificity for stimulating the actin nucleating activity of Arp2/3.<
Figure 1 Legend
Figure 1. Purity determination of GST-WASP VCA domain. 20 ug of VCG03 was run on an SDS-PAGE gel and visualized by coomassie blue staining. Protein quantitation was performed using the Precision Red Protein Assay Reagent (Cat.# ADV02 ).
Figure 2 Legend
Figure 2: Actin polymerization stimulated by Arp2/3 complex and the VCA domain of WASP. Actin polymerization was measured using kit BK003 . The addition of Arp2/3 complex or the VCA domain alone to actin has minimal effects on actin polymerization, while the combination of Arp2/3 and the VCA domain strongly stimulates the rate of actin polymerization.

Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.

All products are intended for research use only (RUO). Not for human, veterinary or therapeutic use.

Amount: 1 x 500 ug
Available: In stock
available

Delivery expected until 1/17/2025 

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