Comparison

Cdc42-His Protein: constituitively active

Item no. C6101-A
Manufacturer Cytoskeleton
Amount 1 x 10 ug
Category
Type Proteins
Specific against other
Purity Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 12% polyacrylamide gel. His-Cdc42(Q61L) protein was determined to be >70% pure.
Citations Zhang, X. F., Schaefer, A. W., Burnette, D. T., Schoonderwoert, V. T. and Forscher, P. (2003). Rho-dependent contractile responses in the neuronal growth cone are independent of classical peripheral retrograde actin flow. Neuron 40, 931-944.
ECLASS 10.1 32160409
ECLASS 11.0 32160409
UNSPSC 12352202
Alias Small G protein, Cdc42 protein, Active cdc42, his-Cdc42
Similar products Small G protein, Cdc42 protein, Active cdc42, his-Cdc42
Shipping Condition Room temperature
Available
Shipping Temperature
AT
Storage Conditions
On Arrival: 4°C
Delivery Time
1-2 Weeks
FAQs

Question 1: Does the constitutively-active mutant Cdc42 protein come pre-loaded with GTP?


Answer 1: The purified small G-proteins that Cytoskeleton provides, whether they are wild-type, constitutively-active or dominant negative proteins, do not come pre-loaded with GTP. Typically, 50% of the protein is bound to GTP and 50% is bound to GDP upon resuspension. Thus, proteins will have to be pre-loaded with either GTP (Cat. #BST06) or GTPgS (Cat. #BS01) for experiments.


Question 2: In what buffer is the mutant Cdc42 protein shipped?


Answer 2: The His-Cdc42 L61 mutant protein ((Cat. #C6101) is supplied as a lyophilized white powder. The protein should be reconstituted to 1 mg/ml by the addition of 10 &mu, l of distilled water. The protein will be in the following buffer: 10 mM Tris pH 7.5, 10 mM NaCl, 0.1 mM MgCl2, 0.5% sucrose and 0.1% dextran. In order to maintain high biological activity of the protein, it is strongly recommended that the protein solution be supplemented with DTT to 1 mM final concentration, aliquoted into "experiment-sized" amounts, snap frozen in liquid nitrogen and stored at -70°C. The protein is stable for 6 months under these conditions. The protein must not be exposed to repeated freeze-thaw cycles.

Weight (grams)
7
Product Uses
Cdc42 effector and GAP identification and binding studies
Activation of Cdc42 effectors in vitro and in vivo
Stimulation of Cdc42 phenotype by microinjection into cells
Material
The constitutively active form of human Cdc42 protein contains a glutamine to leucine substitution at residue 61. The common name for this mutant is Cdc42(Q61L) or L61Cdc42. The leucine substitution prevents endogenous and GAP-stimulated GTPase activity of Cdc42, hence the protein is always in the active, GTP-bound, state. A similar mutation in Ras (Ras(Q61L)) behaves as an oncogenic mutation.
Cdc42(Q61L) has been expressed in a bacterial system. The protein is supplied as a lyophilized powder.When it is reconstituted in distilled water to 1 mg/ml, the protein is in the following buffer: 2 mM Tris pH 7.6, 0.5 mM MgCl2 0.5% sucrose and 0.1% dextran. Protein concentration is determined by the Precision Red Advanced Protein Assay Reagent (Cat. # ADV02).
The recombinant protein is 25 kDa, consisting of the 22 kDa Cdc42 constitutively active protein plus a histidine tag in the amino-terminus.
For other forms of Cdc42 as well as many other purified small G-proteins, see our main small G-protein product page>.
Biological Activity
His-Cdc42(Q61L) mutant protein binds GTP but its intrinsic GTPase activity has been eliminated, resulting in a constitutively active protein. The biological assay for His-Cdc42(Q61L) activity consists of a pulldown assay using PAK-PBD beads (Cat. # PAK02). The PAK protein is an effector of Cdc42 and will specifically bind to active GTP-Cdc42. Stringent quality control ensures that > 80% of His-Cdc42(Q61L) protein can be pulled down using this method.

Figure 1 Legend
His-Cdc42(Q61L) protein purity determination. A 10 ug sample of C6101 (His-Cdc42(Q61L) molecular weight approx. 25 kDa) was separated by electrophoresis in a 12% SDS-PAGE system, and stained with Coomassie Blue.

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 10 ug
Available: In stock
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