Immunofluorescent analysis of Rho A/B/C (green) showing staining in the in the cytoplasm of A431 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Rho A/B/C monoclonal antibody (Product # MA1-011) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Full-length recombinant human Rho A|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||1:500 - 1:1500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Western blot analysis of MA1-011 detects an ~20 kDa protein and shows reactivity to RhoA, RhoB, and RhoC.
RhoA is described as a small GTPase protein that regulates the actin cytoskeleton in the formation of stress fibers. RhoA is also included in the related proteins of the Ras superfamily, which involves proteins in the regulation and timing of cell division.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.