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17-10394 Rac1/cdc42 Activation Magnetic Beads Pulldown Assay

The Rac1/cdc42 Activation Magnetic Beads Pulldown Assay provides an effective method for detecting Rac & Cdc42 activity in cell lysates with higher yield & easier process utilizing magenetic bead properties.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: Affinity Binding Assay, G-Protein Assay, Activity Assay
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17-10394
30 assays  
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      概述

      Replacement Information

      重要规格表

      Species ReactivityKey Applications
      H, M, RABA, GPA, ACT
      Description
      Catalogue Number17-10394
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionRac1/cdc42 Activation Magnetic Beads Pulldown Assay
      OverviewRac1/cdc42 Activation Magnetic Beads Pulldown Assay (Cat. No. 17-10394) provides an effective method for detecting Rac and Cdc42 activity in cell lysates. This assay uses the downstream effector of Rac/Cdc42, p21-activated protein kinase (PAK1), to isolated the active GTP-bound form of Rac/Cdc42 from the sample. The p21 binding domain (PBD) of PAK1 is expressed as a GST-fusion protein and coupled to magnetic beads. After magnetic rack separation, an immunoblot is performed and the activated Rac/Cdc42 is detected with specific monoclonal antibodies, followed by HRP-conjugated secondary antibody and ECL reagent.
      Materials Required but Not Deliveredstimulated and non-stimulated cell lysates pipettes, micropipettes and tips electrophoresis and immunoblotting equipment microcentrifuge microcentrifuge tubes Rac activators glutathione agarose tissue culture supplies 4C tube rocker 30C incubator or water bath glycerol 500 mM EDTA
      References
      Product Information
      Components
      • Rac1/cdc42 Assay Reagent (Pak-1PBD, Magnetic beads): 300 ug of Pak-1 PBD in 150 uL of glutathione magnetic beads, provided as a 50% beads slurry in PBS containing 50% glycerol for a final volume of 300 uL.
      • Anti-Rac1, clone 23A8: One vial containing 250 ug of purified IgG2akappa in 250 uL of storage buffer (0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, containing 0.05% sodium azide).
      • Anti-cdc42, mouse monoclonal IgG1: One vial containing 50 ug of purified mouse IgG1 in 200 uL of 10 mM sodium phosphate, pH 7.5, 75 mM NaCl, 0.75 mM sodium azide, 0.5 mg/mL BSA and 50% glycerol.
      • Mg2+ Lysis/Wash Buffer, 5X, Catalog # 20-168 : Two vials, each vial containing 18 mL of 5X MLB: 125 mM HEPES, pH 7.5, 750 mM NaCl, 5% Igepal CA-630, 50 mM MgCl2, 5 mM EDTA and 10% glycerol.
      • 100X GTPγS, 10mM, Catalog # 20-176: One vial containing 50 μL of 10 mM GTPγS, 100X stock, in 50 mM Tris-HCl, pH 7.8, non-hydrolyzable analog of GTP. Sufficient to label 5 mL of cell lysates.
      • 100X GDP, 100mM, Catalog # 20-177: One vial containing 50 μL of 100 mM GDP, 100X stock, in 50 mM Tris-HCl, pH 7.8. GDP (Guanosine 5'-Diphosphate) for in vitro labeling of G-proteins in the inactive form. Sufficient to label 5ml of cell lysates.
      Quality LevelMQ100
      Applications
      ApplicationThe Rac1/cdc42 Activation Magnetic Beads Pulldown Assay provides an effective method for detecting Rac & Cdc42 activity in cell lysates with higher yield & easier process utilizing magenetic bead properties.
      Key Applications
      • Affinity Binding Assay
      • G-Protein Assay
      • Activity Assay
      Biological Information
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Species Reactivity NoteRac/cdc42 Assay Reagent (PAK-1 PBD, magnetic beads): Species Cross-reactivity: Human and mouse. Predicted to cross-react with all mammalian species.
      Anti-Rac1, clone 23A8: Species Cross-reactivity: Human, mouse and rat. Other species cross-reactivity unknown.
      Anti-cdc42 Species Cross-reactivity: Human, mouse, rat and dog. Other species cross-reactivity unknown.
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene is a GTPase which belongs to the RAS superfamily of small GTP-binding proteins. Members of this superfamily appear to regulate a diverse array of cellular events, including the control of cell growth, cytoskeletal reorganization, and the activation of protein kinases. Several alternatively spliced transcript variants of this gene have been described, but the full-length nature of some of these variants has not been determined.
      Gene Symbol
      • RAC1
      • MGC111543
      • ORFNames=MIG5
      • PAK1
      • MIG5
      • TC-25
      • p21-Rac1
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P63000 # Isoform B has an accelerated GEF-independent GDP/GTP exchange and an impaired GTP hydrolysis, which is restored partially by GTPase-activating proteins. It is able to bind to the GTPase-binding domain of PAK but not full-length PAK in a GTP- dependent manner, suggesting that the insertion does not completely abolish effector interaction.
      SIZE: 192 amino acids; 21450 Da
      SUBUNIT: Interacts with the GEF proteins PREX1, RASGRF2, DOCK1, DOCK2 and DOCK7, which promote the exchange between GDP and GTP, and therefore activate it. Interacts with PARD6A, PARD6B and PARD6G in a GTP-dependent manner. Part of a quaternary complex containing PARD3, some PARD6 protein (PARD6A, PARD6B or PARD6G) and some atypical PKC protein (PRKCI or PRKCZ), which plays a central role in epithelial cell polarization. Found in a trimeric complex composed of DOCK1 and ELMO1, which plays a central role in phagocytosis of apoptotic cells. Interacts with RALBP1 via its effector domain. Interacts with PLXNB1. Part of a complex with MAP2K3, MAP3K3, CCM2 and DEF6. Interacts with BAIAP2, CYFIP1/SRA-1 and DEF6. Interacts with Y.pseudotuberculosis YPKA and PLCB2.
      SUBCELLULAR LOCATION: Cell membrane; Lipid-anchor; Cytoplasmic side (By similarity). Melanosome. Note=Inner surface of plasma membrane possibly with attachment requiring prenylation of the C- terminal cysteine (By similarity). Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
      TISSUE SPECIFICITY: Isoform B is predominantly identified in skin and epithelial tissues from the intestinal tract. The expression of isoform B is elevated in colorectal tumors at various stages of neoplastic progression, as compared to their respective adjacent tissues.
      DOMAIN: SwissProt: P63000 The effector region mediates interaction with DEF6.
      SIMILARITY: Belongs to the small GTPase superfamily. Rho family.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely evaluated by precipitating Rac1-GTP and cdc42-GTP from 3T3/A31 and HeLa cell lysates that had been loaded with GTPγS(lysates were loaded in vitro with GTPγS for cdc42-GTP). The precipitated Rac1-GTP and cdc42-GTP was detected by immunoblot analysis using anti-Rac1 (1 μg/ml).
      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.
      Storage and Shipping Information
      Storage Conditions1 year at -20°C from date of shipment
      Packaging Information
      Material Size30 assays
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      产品目录编号 GTIN
      17-10394 04053252930980

      相关产品和应用

      Accessories

      产品目录编号 描述  
      09-258 Anti-phospho-PAK1 (Ser199/Ser204) Antibody 显示价格及库存情况
      09-271 Anti-Rac1b Antibody 显示价格及库存情况

      Kit Component

      产品目录编号 描述  
      05-389 Anti-Rac1 Antibody, clone 23A8 显示价格及库存情况
      20-168 Mg2+ Lysis/Wash Buffer, 5X 显示价格及库存情况
      20-176 100X GTPγS, 10mM 显示价格及库存情况
      20-177 100X GDP, 100mM 显示价格及库存情况

      种类

      Life Science Research > Protein Detection and Quantification > Activity Assays > Activation Assays