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17-10393 Rac1 Activation Magnetic Beads Pulldown Assay

17-10393
30 assays  Kit capacity: 30 assays
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

Species ReactivityKey Applications
H, M, RABA
Description
Catalogue Number17-10393
ReplacesSGT425
Brand Family Upstate
Trade Name
  • Upstate
DescriptionRac1 Activation Magnetic Beads Pulldown Assay
References
Product Information
Components
  • Rac1/Cdc42 Assay Reagent (Pak-1 PBD, 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: (Part No. 05-389) 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).
  • Mg2+ Lysis/Wash Buffer, 5X, (Part No. 20-168).Two vials, each vial contains 18 mL of 125 mM, HEPES, pH 7.5, 750 mM NaCl, 5% Igepal CA-630, 50mM MgCl2, 5 mM EDTA and 10% glycerol
  • 100X GTPγS, 10mM (Cat.# 20-176) One vial containing 50 uL of 10 mM GTPγS, 100X stock, in sterile water. Non-hydrolyzable analog of GTP. Sufficient to label 5 mL of cell lysates
  • 100X GDP, 100 mM, (Part No. 20-177). One vial containing 50 uL of 100 mM GDP, 100X stock, in sterile water. GDP, Guanosine 5’-Diphosphate, type I, for in vitro labeling of G-proteins in the inactive form. Sufficient to label 5ml of cell lysates.
Quality LevelMQ100
Applications
ApplicationNon-radioactive Rac1 Activation Assay Kit can be used to precipitate Rac1-GTP from cell lysates, & detection by a Rac1 specific monoclonal antibody.
Key Applications
  • Affinity Binding Assay
Biological Information
Species Reactivity
  • Human
  • Mouse
  • Rat
Species Reactivity NoteRac1/Cdc42 Assay Reagent (PAK-1 PBD, magnetic beads): Human and mouse. Predicted to cross-react with all mammalian species
Anti-Rac1, clone 23A8 (mouse monoclonal IgG2b): Human, mouse and rat, other species cross-reactivity unknown. Note: This antibody may be reused 3-5 times for immunoblot analysis. Add 0.05% sodium azide to the blocking buffer containing the diluted antibody. Do not use sodium azides with HRP
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 Rac-GTP from 3T3 cell lysates that had been loaded with GTPγS. The precipitated Rac-GTP was detected by immunoblot analysis using anti-Rac.
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
Packaging Information
Material Size30 assays
Material PackageKit capacity: 30 assays
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
17-10393 04053252930973