手机验证
Cdc42 Pulldown Activation Assay Kit
研卉生物
20 assays
Product Uses Include
Introduction
The Rho switch operates by alternating between an active, GTP-bound state and an inactive, GDP-bound state. Understanding the mechanisms that regulate activation / inactivation of the GTPases is of obvious biological significance and is a subject of intense investigation. The fact that many Rho family effector proteins will specifically recognize the GTP bound form of the protein has been exploited experimentally to develop a powerful affinity purification assay that monitors Rac and Cdc42 protein activation. The assay uses the Cdc42/Rac Interactive Binding (CRIB) region (also called the p21 Binding Domain, PBD) of the Cdc42 / Rac effector protein, p21 activated kinase 1 (PAK). The CRIB/PBD protein motif has been shown to bind specifically to the GTP-bound form of Rac and/or Cdc42 proteins. The fact that the PBD region of PAK has a high affinity for both GTP-Rac and GTP-Cdc42 and that PAK binding results in a significantly reduced intrinsic and catalytic rate of hydrolysis of both Rac and Cdc42 make it an ideal tool for affinity purification of GTP-Rac and GTP-Cdc42 from cell lysates. The PAK-PBD protein supplied in this kit corresponds to residues 67-150. This includes the highly conserved CRIB region (aa 74-88) plus sequences required for the high affinity interaction with GTP-Rac and GTP-Cdc42. The PAK-PBD is in the form of a GST fusion protein, which allows one to "pull-down" the PAK-PBD/GTP-Cdc42 (or GTP-Rac) complex with glutathione affinity beads. The assay therefore provides a simple means of quantitating Rac/Cdc42 activation in cells. The amount of activated Cdc42 is determined by a Western blot using a Cdc42 specific antibody.
Kit contents
The kit contains sufficient materials for 20 assays, depending on assay setup, and includes reagents for positive and negative controls. A larger 50 assay version of this kit is available as Cat. # BK034. The following components are included:
|
||
|
Figure 1. The brightly colored glutathione agarose beads in BK034-S makes the kit easy to use. |
Equipment needed
Example results
The Cdc42 activation assay was tested by loading the Cdc42 protein in cell lysates with either GTPγS or GDP. As expected, the GTPγS-loaded Cdc42 is very efficiently precipitated while very little GDP-loaded Cdc42 is precipitated (Fig. 2).
|
||
|
Figure 2. Results from BK034-S Cdc42 activation assay. Activated Cdc42 was precipitated and detected in a Western blot using kit BK034-S. The first lane shows a 50 ng recombinant His-tagged Cdc42 standard (Rec. His-Cdc42). The following lanes shows the pull-down of inactive, GDP-loaded Cdc42 (Cdc42-GDP PD) or active, GTPγS-loaded Cdc42 (Cdc42-GTP PD) from equal amounts of cell lysates. |
Sakabe et al., 2012. Age-related guanine nucleotide exchange factor, mouse Zizimin2, induces filopodia in bone marrow-derived dendritic cells. Immun. Ageing. v 9, p 2.
Nithipatikom et al., 2012. Cannabinoid Receptor Type 1 (CB1) Activation Inhibits Small GTPase RhoA Activity and Regulates Motility of Prostate Carcinoma Cells. Endocrinology. v 153, pp 29-41.
Asai et al., 2011. Involvement of Rac GTPase activation in phosphatidylcholine hydroperoxide-induced THP-1 cell adhesion to ICAM-1. Biochem. Biophys. Res. Commun. v 406, pp 273-277.
Abu-Elneel et al., 2008. A delta-Catenin Signaling Pathway Leading to dendritic protrusions. J. Biol. Chem. v 283, pp 32781-32791.
Nur-E-Kamal et al., 2005. Three dimensional nanofibrillar surfaces induce activation of Rac. Biochem. Biophys. Res. Commun. v 331, pp 428-434.
Slice et al., 2005. Angiotensin II and epidermal growth factor induce cyclooxygenase-2 expression in intestinal epithelial cells through small GTPases using distinct signaling pathways. J. Biol. Chem. v 280, pp 1582-1593.
Tang et al., 2005. The Adapter Protein CrkII Regulates Neuronal Wiskott-Aldrich Syndrome Protein, Actin Polymerization, and Tension Development during Contractile Stimulation of Smooth Muscle. J. Biol. Chem. v 280, pp 23380-23389.
Liu et al., 2004. Nucleotide exchange factor ECT2 interacts with the polarity protein complex Par6/Par3/protein kinase Cζ (PKCζ) and regulates PKCζ activity. Mol. Cell. Biol. v 24, pp 6665-6675.
Sasai et al., 2004. The neurotrophin-receptor-related protein NRH1 is essential for convergent extension movements. Nat. Cell Biol. v 6, pp 741-748.
Tang and Gunst, 2004. The small GTPase Cdc42 regulates actin polymerization and tension development during contractile stimulation of smooth muscle. J. Biol. Chem. v 279, pp 51722-51728.
Cytoskeleton | BK030 | Combo RhoA/Rac1/Cdc42 Activation Assay Kit | KIT 3 x 10 assays |
Cytoskeleton | BK032-S | Arf1 Pulldown Activation Assay Kit | KIT 20 assays |
Cytoskeleton | BK033-S | Arf6 Pulldown Activation Assay Kit | KIT 20 assays |
Cytoskeleton | BK034 | Cdc42 Pulldown Activation Assay Kit | KIT 50 assays |
Cytoskeleton | BK034-S | Cdc42 Pulldown Activation Assay Kit | KIT 20 assays |
Cytoskeleton | BK035 | Rac1 Pulldown Activation Assay Kit | KIT 50 assays |
Cytoskeleton | BK035-S | Rac1 Pulldown Activation Assay Kit | KIT 20 assays |
Cytoskeleton | BK036 | RhoA Pulldown Activation Assay Kit | KIT 80 assays |
Cytoskeleton | BK036-S | RhoA Pulldown Activation Assay Kit | KIT 20 assays |
Cytoskeleton | BK037 | G-Actin : F-Actin In Vivo Assay Kit | KIT 30-100 assays |
Cytoskeleton | BK038 | Microtubule / Tubulin In Vivo Assay Kit | KIT 30-100 assays |
Cytoskeleton | BK040 | RalA Activation Assay Biochem Kit | KIT 50 assays |
Cytoskeleton | BK051/52 | ATPase ELIPA™ (Enzyme Linked Inorganic Phosphate Assay) | KIT 96 assays |
Cytoskeleton | BK051/52 | GTPase ELIPA™ (Enzyme Linked Inorganic Phosphate Assay) | KIT 96 assays |
Cytoskeleton | BK053 | HTS Kinesin ATPase Endpoint Assay | KIT 1000 assays |
Cytoskeleton | BK054 | CytoPhos™ Phosphate Assay (1-500ug/ml protein reactions) | KIT 1000 assays |
Cytoskeleton | BK060 | Kinesin ELIPA (Enzyme Linked Inorganic Phosphate Assay TM) | KIT 96 assays |
Cytoskeleton | BK100 | GEF Exchange Assay | KIT 60-300 assays |
Cytoskeleton | BK105 | GAP Assay Kit | KIT 80-160 assays |
Cytoskeleton | BK121 | RhoA G-LISA Activation Assay (luminescence) | KIT 96 assays |
Cytoskeleton | BK124-S | RhoA G-LISA Activation Assay (absorbance) | KIT 24 assays |
Cytoskeleton | BK124 | RhoA G-LISA Activation Assay (absorbance) | KIT 96 assays |
Cytoskeleton | BK125 | RhoA G-LISA Activation Assay (absorbance) | Large Quantities |
Cytoskeleton | BK125 | Rac1,2,3 G-LISA Activation Assay Kit | KIT 96 assays |
Cytoskeleton | BK126 | Rac1 G-LISA Activation Assay Kit | KIT 96 assays |
Cytoskeleton | BK127 | Cdc42 G-LISA Activation Assay Kit | KIT 96 assays |
Cytoskeleton | BK127-S | Cdc42 G-LISA Activation Assay Kit | KIT 24 assays |
Cytoskeleton | BK128 | Rac1 G-LISA Activation Assay Kit | KIT 96 assays |
Cytoskeleton | BK128-S | Rac1 G-LISA Activation Assay Kit | KIT 24 assays |
Cytoskeleton | BK129 | RalA G-LISA Activation Assay Kit | KIT 96 assays |
Cytoskeleton | BK131 | Ras G-LISA Activation Assay Kit | KIT 96 assays |
Cytoskeleton | BK132 | Ras G-LISA Activation Assay Kit | Large Quantities |
Cytoskeleton | BK132 | Arf1 G-LISA Activation Assay Kit | KIT 96 assays |
Cytoskeleton | BK133 | Arf6 G-LISA Activation Assay Kit | KIT 96 assays |
Cytoskeleton | BK135 | G-LISA Bundle (BK135 = BK124-S + BK127-S + BK128-S) | 3 KITS (24 assays/kit) |
Cytoskeleton | BK150 | RhoA ELISA Kit | KIT 96 assays |
Cytoskeleton | BK160 | Signal-Seeker™ Phosphotyrosine Enrichment Kit (30 assays) | KIT 30 assays |
Cytoskeleton | BK161 | Signal-Seeker™ Ubiquitin Enrichment Kit (30 assays) | KIT 30 assays |
Cytoskeleton | BK162 | Signal-Seeker™ SUMOylation Enrichment Kit (30 assays) | KIT 30 assays |
风险提示:丁香通仅作为第三方平台,为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全,合理判断,谨慎选购商品,商家和用户对交易行为负责。对于医疗器械类产品,请先查证核实企业经营资质和医疗器械产品注册证情况。