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.
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About This Item
Código UNSPSC:
12161503
eCl@ss:
32161000
NACRES:
NA.32
Productos recomendados
Nivel de calidad
reactividad de especies
human, rat, mouse
fabricante / nombre comercial
Upstate®
técnicas
activity assay: suitable
affinity binding assay: suitable (G-protein)
Nº de acceso NCBI
Nº de acceso UniProt
Condiciones de envío
dry ice
Información sobre el gen
human ... RAC1(5879)
Descripción general
Rac1/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.
Especificidad
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.
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.
Aplicación
Research Category
Cell Signaling
Signaling
Cell Signaling
Signaling
Componentes
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.
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.
Calidad
Routinely 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).
Almacenamiento y estabilidad
1 year at -20°C from date of shipment
Información legal
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Cláusula de descargo de responsabilidad
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.
Palabra de señalización
Danger
Frases de peligro
Consejos de prudencia
Clasificaciones de peligro
Aquatic Acute 1 - Aquatic Chronic 2 - Eye Dam. 1
Código de clase de almacenamiento
10 - Combustible liquids
Certificados de análisis (COA)
Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»
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