Anti-Phospho-ROCK2 (Tyr256) Polyclonal Antibody is a Rabbit antibody targeting Phospho-ROCK2 (Tyr256). Anti-Phospho-ROCK2 (Tyr256) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog,Pig,Cow,GuineaPig)

1. Tissue/cell: rat testis tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-phospho-ROCK2 (Tyr256) Polyclonal Antibody, Unconjugated (TMAB-11239) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Paraformaldehyde-fixed, paraffin embedded (Mouse testis); 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-ROCK2 (Tyr256)) Polyclonal Antibody, Unconjugated (TMAB-11239) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-phospho-ROCK2 (Tyr256) antibody (TMAB-11239)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at 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.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 0.2μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human ROCK2 around the phosphorylation site of Tyr256

Human

ROCK2

Rho-associated protein kinase 2

Regulates the assembly of the actin cytoskeleton. Promotes formation of stress fibers and of focal adhesion complexes. Plays a role in smooth muscle contraction.