Anti-Phospho-RhoA (Ser188) Polyclonal Antibody is a Rabbit antibody targeting Phospho-RhoA (Ser188). Anti-Phospho-RhoA (Ser188) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Rat (predicted:Mouse,Chicken,Dog,Cow,Sheep)

1. Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Phospho-Rhoa (Ser188)) Polyclonal Antibody, Unconjugated (TMAB-01488) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Sample: Brain (Rat) Lysate at 40 μg
Primary: Anti-Phospho-Rhoa (Ser188) (TMAB-01488) at 1/300 dilution
Secondary: HRP conjugated Goat-Anti-rabbit IgG (secondary antibody) at 1/5000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDa
3. Blank control (Black line): Molt4 (Black). Primary Antibody (green line): Rabbit Anti-Phospho-RhoA (Ser188) antibody Dilution: 3 μg/10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647 Dilution: 3 μg/test. Protocol The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
4. Molt-4 cells were fixed with 4% PFA for 10 min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Phospho-RhoA Antibody (TMAB-01488) at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human RhoA around the phosphorylation site of Ser188

Human

RHOA

Transforming protein RhoA

Ras Family,Actin Assembly,Ras family

Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Involved in a microtubule-dependent signal that is required for the myosin contractile ring formation during cell cycle cytokinesis. Plays an essential role in cleavage furrow formation. Required for the apical junction formation of keratinocyte cell-cell adhesion. Serves as a target for the yopT cysteine peptidase from Yersinia pestis, vector of the plague, and Yersinia pseudotuberculosis, which causes gastrointestinal disorders. Stimulates PKN2 kinase activity. May be an activator of PLCE1. Activated by ARHGEF2, which promotes the exchange of GDP for GTP. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. The MEMO1-RHOA-DIAPH1 signaling pathway plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. It controls the localization of APC and CLASP2 to the cell membrane, via the regulation of GSK3B activity. In turn, membrane-bound APC allows the localization of the MACF1 to the cell membrane, which is required for microtubule capture and stabilization.