Phospho-RhoA (Ser188) Rabbit Polyclonal Antibody

• Catalog Number: orb6455
• Category: Antibodies
• Description: Phospho-RhoA (Ser188) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Gallus, Mouse, Sheep
• Reactivity: Human, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human RhoA around the phosphorylation site of Ser188 KK(p-S)GC
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 21 kDa
• Target: RHOA
• UniProt ID: P61586
• RRID: AB_10933502
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: RHOA(phospho S188); RhoA(phospho S188); RhoA prote
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Gu, Ying et al. Fasudil attenuates soluble fms-like tyrosine kinase-1 (sFlt-1)-induced hypertension in pregnant mice through RhoA/ROCK pathway Oncotarget, 8, 104104-104112 (2017)

PubMed: 29262624

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. Acquisition of 20000 events was performed.

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 (orb6455) 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 20000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Phospho-RhoA (Ser188)) Polyclonal Antibody, Unconjugated (orb6455) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Brain (Rat) Lysate at 40 ug, Primary: Anti-Phospho-RhoA (Ser188) (orb6455) at 1/300 dilution, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1/5000 dilution, Predicted band size: 21 kD, Observed band size: 21 kD.