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17-10394

Sigma-Aldrich

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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eCl@ss:
32161000
NACRES:
NA.32

Quality Level

species reactivity

human, rat, mouse

manufacturer/tradename

Upstate®

technique(s)

activity assay: suitable
affinity binding assay: suitable (G-protein)

UniProt accession no.

shipped in

dry ice

Gene Information

human ... RAC1(5879)

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This Item
17-44117-28317-286
vibrant-m

17-441

Rac1/Cdc42 Activation Assay Kit

vibrant-m

17-283

Rac1 Activation Assay Kit

vibrant-m

17-286

Cdc42 Activation Assay Kit

species reactivity

human, rat, mouse

species reactivity

human, mouse, rat

species reactivity

human, rat, mouse

species reactivity

-

Quality Level

100

Quality Level

100

Quality Level

100

Quality Level

100

manufacturer/tradename

Upstate®

manufacturer/tradename

Upstate®

manufacturer/tradename

Upstate®

manufacturer/tradename

Upstate®

technique(s)

activity assay: suitable, affinity binding assay: suitable (G-protein)

technique(s)

activity assay: suitable, affinity binding assay: suitable (G-protein)

technique(s)

activity assay: suitable, affinity binding assay: suitable

technique(s)

activity assay: suitable, affinity binding assay: suitable

NCBI accession no.

NM_018890.2, NM_006908.3, NM_198829.1

NCBI accession no.

NM_006908.3, NM_018890.2, NM_198829.1

NCBI accession no.

NM_006908.3, NM_018890.2, NM_198829.1

NCBI accession no.

NM_001039802.1, NM_001791.3, NM_044472.2

General description

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.

Specificity

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.

Application

Research Category
Cell Signaling

Signaling

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

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).

Storage and Stability

1 year at -20°C from date of shipment

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

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.

pictograms

CorrosionEnvironment

signalword

Danger

Hazard Classifications

Aquatic Acute 1 - Aquatic Chronic 2 - Eye Dam. 1

Storage Class

10 - Combustible liquids

wgk_germany

WGK 3


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