bs-5330R [Primary Antibody]
Rabbit  Anti-Phospho-RhoA (Ser188) Rabbit pAb  Polyclonal Antibody
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Host: Rabbit

Target Protein: Phospho-RhoA (Ser188) Rabbit pAb

IR: Immunogen Range:KK(p-S)GC

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 387

Swiss Prot: P61586

Source: KLH conjugated Synthesised phosphopeptide derived from human RhoA around the phosphorylation site of Ser188:KK(p-S)GC 

Purification: affinity purified by Protein A

Storage: 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.

Background: This gene encodes a member of the Rho family of small GTPases, which cycle between inactive GDP-bound and active GTP-bound states and function as molecular switches in signal transduction cascades. Rho proteins promote reorganization of the actin cytoskeleton and regulate cell shape, attachment, and motility. The protein encoded by this gene is prenylated at its C-terminus, and localizes to the cytoplasm and plasma membrane. It is thought to be important in cell locomotion. Overexpression of this gene is associated with tumor cell proliferation and metastasis. Multiple alternatively spliced variants, encoding the same protein, have been identified.

Size: 200ul

Concentration: 1mg/ml

Applications: WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1μg/Test

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

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Sample: Brain (Rat) Lysate at 40 ug Primary: Anti-Phospho-RhoA(Ser188) (bs-5330R) at 1/300 dilution Secondary: HRP conjugated Goat-Anti-rabbit IgG (bs-0295G-HRP) at 1/5000 dilution Predicted band size: 21 kD Observed band size: 21 kD
Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-RhoA(Ser188)) Polyclonal Antibody, Unconjugated (bs-5330R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Molt-4 cells were fixed with 4% PFA for 10min 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(bs-5330R)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).
Blank control (Black line): Molt4 (Black). Primary Antibody (green line):Rabbit Anti-Phospho-RhoA (Ser188) antibody (bs- 5330R) Dilution: 3μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647 Dilution: 3μg /test. Protocol The cells were fixed with 4% PFA (10min 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 20,000 events was performed.
 
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