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Description
Pulldown based small G-protein activation assays work by using a domain of an effector protein coupled to agarose beads. These affinity beads only bind the active (GTP-bound) form of the protein, thereby isolating the active pool of target small g-protein from a complex cell or tissue homgenate. The isolated material is run on a Western blot which is probed with an antibody to a specific isotype e.g. RhoA. Cytoskeleton's kits have many more reagents than other suppliers (see Contents section Manual in Documents tab), which allows you non-stop experimental operation at every point in the procedure. In addition we provide affinity beads alone for greater flexibility in experimental design, PAK-RBD for Rac and Cdc42 (Cat. # PAK02), and Rhotekin-RBD for Rho (Cat. # RT02). An image of the beads is shown to the left, you will notice the blue colored beads which allow easy identificaiton of the pellet during the centrifuge based washing procedure. Cytoskeleton was first to introduce colored beads in 2004 in response to customers requests to better visualize the pellet.
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Pull-down Assays
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