• Combo kit for detection of activated (GTP-bound) Cdc42/Rac1 and RhoA protein in cell and tissue lysates
• Compare Cdc42, Rac1 and RhoA signaling pathways

Kit contents (3 x 10 assays)

• Rhotekin-PBD beads for affinity purification of active RhoA, B & C
• RhoA-specific antibody to detect activated RhoA only
• His-RhoA control protein
• P21 activated kinase1 protein (PAK)-PBD beads for affinity purification of active Cdc42 & Rac
• Cdc42-specific antibody to detect activated Cdc42 only
• His-Cdc42 control protein
• Rac1-specific antibody to detect activated Rac1 only
• His-Rac1 control protein
• Cell lysis buffer-optimized for maintenance of GTP-bound GTPases
• Wash buffer-optimized for maintenance of GTP-bound GTPases
• Loading buffer: for control reactions
• STOP buffer: for control reactions
• GTPγS: for control reactions
• GDP: for control reactions
• Protease inhibitor cocktail
• DMSO: for protease resuspension

Equipment & materials required

• Cell lysates
• Cell scraper for harvesting cells
• Liquid nitrogen for snap freezing lysates
• Equipment for Western blot performance & analysis
• Microcentrifuge

The BK030 kit is a combination sampler size of the Rac1, Cdc42 and RhoA pull-down kits. There is sufficient reagent to perform 10 pull-down assays for each GTPase.

Key characteristics

• Semi-quantitative assay
• Detection sensitivity: See BK035, BK034 & BK036 specifications
• Detection specificity: Rac1, Cdc42 & RhoA proteins
• Input requirements: 200 µg to 1mg of cell or tissue lysate per assay
• Timeframe: Approximately 10 hours, typically performed over two days