All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: ARPD-50 (for 50T)

Cat. No.: ARPD-250 (for 250T)

Description

Active Rho Pull-down and Detection Kit, also known as Active Rho Binding Precipitation and Detection Reagent Kit, is a convenient and efficient kit used for quantitatively detecting activated state Rho proteins (including RhoA, RhoB, and RhoC) in tissue or cell lysates. This kit is based on the specific precipitation (pull-down) of activated state Rho proteins using Rhotekin-RBD Agarose (active Rho binding agarose gel), followed by Western detection to assess the activation level of Rho proteins.

Rhotekin is an effector protein that interacts with Rho proteins, including RhoA, RhoB, and RhoC, regulating various cellular activities such as proliferation of neuronal precursor cells, growth of neuronal processes, and synaptic formation. The term "teki" in Rhotekin comes from the Japanese word for "target." Rhotekin consists of 563 amino acids with a theoretical molecular weight of 61 kDa and contains four domains: the active Rho-binding domain (RBD), pleckstrin homology (PH) domain, and two proline-rich motif domains. The RBD domain of Rhotekin specifically recognizes and binds to activated state Rho proteins but not to inactive state Rho proteins. Therefore, recombinant Rhotekin-RBD protein can be used to detect activated state Rho proteins and can also serve as an inhibitor of Rho proteins in vivo or in vitro.

Small GTPases, also known as Small G-proteins or the Ras superfamily, are molecular switches that regulate processes such as signal transduction, cell proliferation, cytoskeletal reorganization, and intracellular membrane trafficking in eukaryotic cells. Small GTPases cycle between an "active" and an "inactive" state by binding and hydrolyzing GTP. Upon stimulation by extracellular signals, guanine nucleotide-exchange factors (GEFs) assist small GTPases in exchanging bound GDP for GTP, leading to the activation of the small GTPase. Activated small GTPases interact with effector proteins downstream, stimulating cellular responses accordingly. GTPase-activating proteins (GAPs) catalyze the hydrolysis of GTP bound to small GTPases into GDP and release free phosphate (Pi), causing the small GTPase to enter an inactive state. Guanine nucleotide dissociation inhibitors (GDIs) inhibit the release of GDP from the small GTPase until GEFs receive stimulation signals to initiate a new cycle.

Components

• Rhotekin-RBD Agarose
• Recombinant Human RhoA (10ng/μl)
• RhoA Rabbit Monoclonal Antibody (1000X)
• GppNHp (100X)
• GDP (100X)
• Cell Lysis Buffer
• Wash Buffer
• 100mM EDTA (pH8.0)
• 60mM MgCl₂
• Protease Inhibitor Cocktail (100X)

Storage

Stored at -80°C, valid for one year. Except for Rhotekin-RBD Agarose, stored at -20°C, valid for one year. Rhotekin-RBD Agarose, valid for up to 3 months when stored at -20°C, and up to 1 month when stored at 4°C. Cell Lysis Buffer, Wash Buffer, 100mM EDTA (pH8.0), and 60mM MgCl₂ are valid for up to 3 months when stored at 4°C.

Precautions

• Upon receipt, it is recommended to store Rhotekin-RBD Agarose at -80°C. It is stable for two freeze-thaw cycles, but repeated freeze-thaw should be avoided. For the first use, it is advisable to aliquot appropriately. Other reagents can be stored at -20°C.
• The number of uses of this kit is based on using 20μl or 50μl of Rhotekin-RBD Agarose per assay. When using different amounts of Rhotekin-RBD Agarose, the amounts of other reagents need to be adjusted proportionally.
• This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, food, or drugs. It should not be stored in residential areas.
• For your safety and health, please wear laboratory attire and disposable gloves while handling.

Related:

• Active Rho Pull-down and Detection Kit
• Recombinant Human RhoA (Flag-Tag)
• Rhotekin-RBD Agarose

Only for research and not intended for treatment of humans or animals

Journals Using SBS Genetech Products                                       Universities Using SBS Genetech Products

SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory