G-LISA Bundle (BK135 = BK124-S + BK127-S + BK128-S) (colorim - 3 KITS (24 assays/kit)

Not sure which small G-protein is activated? Our cost-effective combo pull-down kit includes all necessary reagents for 10 assays per GTPase, perfect for identifying your pathway of interest.

Applications

• Combo kit for detection of activated (GTP-bound) Cdc42, Rac1 and RhoA protein in cell and tissue lysates.
• Assay of choice when lysate quantity is limiting, e.g., primary cell cultures
• Compare Cdc42, Rac1 and RhoA signaling pathways

Material

Kit contents (3 x 24 assays: strip-wells allow single or multiple assays per run)

• 24 well Cdc42-GTP binding plate
• 24 well Rac1-GTP binding plate
• 24-well RhoA-GTP binding plate
• Anti Rac1 antibody
• Anti Cdc42 antibody
• Anti-RhoA antibody
• Cdc42 control protein (constitutively active Cdc42)
• Rac1 control protein (constitutively active Rac1)
• RhoA control protein (constitutively active RhoA)
• Secondary detection HRP labeled antibody
• Cell lysis buffer- optimized for maintenance of GTP-bound GTPases
• Binding buffer
• Wash buffer
• Antigen-presenting buffer
• Antibody dilution buffer
• HRP colorimetric detection reagents A & B
• HRP stop solution
• Precision Red advanced protein assay reagent
• Protease inhibitor cocktail
• Strip holder

Equipment & materials required

• Cell lysates
• Cell scraper for harvesting cells
• Concentrated sulfuric acid (1 ml)
• Liquid nitrogen for snap freezing lysates
• Orbital microplate shakers (min 200 rpm) one at 4°C and one at room temperature
• Multiwell pipette
• Microplate spectrophotometer (490 nm)

Biological Activity Assay

The BK135 kit is a combination sampler size of the Rac1, Cdc42 and RhoA colorimetric G-LISA kits. There is sufficient reagent to perform 24 G-LISA assays for each GTPase.

Key characteristics

• Quantitative assay
• Detection sensitivity: See BK128, BK127 & BK124 specifications
• Detection specificity: Rac1, Cdc42 & RhoA proteins
• Input requirements: 10 – 50 µg of cell or tissue lysate per assay
• Timeframe: Approximately 3 hours