The Rho family (Ras homologous GTPases) is highly conserved in eukaryotes and is a key factor in the regulation of actin reorganization. The Rho activity assay can be used to study cell migration, wound healing, cell adhesion, cell polarity, membrane translocation, and cytokinesis. Rho Pull-Down selectively enriches and detects GTP-bound Rho GTPase by activating specific interactions between the GTPase and Rhotekin protein binding domains. The Rho Pull-Down assay can be used to track Rho GTPase activity during cell differentiation, migration, division and cytoskeletal rearrangement.
Figure 1. Schematic diagram of the cycling of small GTPases between "active" and "inactive" states.
Beyotime Active Rho Pull-down and Detection Kit (P2065S/M) allows for convenient and efficient quantitative assay for active Rho proteins (including RhoA, RhoB, and RhoC) in tissue or cell lysates. This kit utilizes Rhotekin-RBD Agarose to specifically bind and precipitate (pull-down) active Rho proteins, and then detect the activation level of Rho proteins through Western blot. The experimental results have been validated using NIH 3T3 cell lysates.
For other related experiments, Beyotime has also launched Rhotekin-RBD Agarose (P2061) and Recombinant Human RhoA (Flag-Tag) (P2063).
Working Principle:
Figure 2. Schematic diagram of the pull-down principle of Beyotime Active Rho Pull-down and Detection Kit (P2065).
Typical Figures:
Figure 3. The assay effect of Beyotime Active Rho Pull-down and Detection Kit (P2065) on endogenous active RhoA protein in NIH 3T3 cells.
Figure 4. The Pull-down effect of Beyotime Rhotekin-RBD Agarose (活性Rho结合琼脂糖凝胶) (P2061).
Figure 5. The SDS-PAGE result of Beyotime Recombinant Human RhoA (P2063).
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