Phospho-ROCK1 (Thr455+Ser456) Rabbit Polyclonal Antibody

• Catalog Number: orb100140
• Category: Antibodies
• Description: Phospho-ROCK1 (Thr455+Ser456) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Canine, Equine, Porcine, Rabbit, Rat
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated synthesised phosphopeptide derived from human ROCK1 around the phosphorylation site of Thr455+Ser456 CR(p-T)(p-S)N
• 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=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 158 kDa
• Target: ROCK1
• UniProt ID: Q13464
• 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: p-ROCK1(Thr455/Ser456); p160 ROCK1; p160ROCK; Rena
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Andalib, Mohammad Nahid et al. The role of RhoA kinase (ROCK) in cell alignment on nanofibers Acta Biomater, 9, 7737-7745 (2013)

PubMed: 23587628

Blank control (blue): Hela (fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody: Rabbit Anti-phospho-ROCK1 (Thr455+Ser456) antibody (orb100140), Dilution: 1 µg in 100 µL 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions), Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.

Blank control: MCF-7. Primary Antibody (green line): Rabbit Anti-phospho-ROCK1 (Thr455+Ser456) antibody (orb100140), Dilution: 1 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, Dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, Protocol, The cells were 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.

Blank control: MCF-7. Primary Antibody (green line): Rabbit Anti-phospho-ROCK1 (Thr455+Ser456) antibody (orb100140), Dilution: 1 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, Dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, Protocol, The cells were 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.

Paraformaldehyde-fixed, paraffin embedded (Human kidney), 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-ROCK1 (Thr455+Ser456)) Polyclonal Antibody, Unconjugated (orb100140) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Cerebrum (Mouse) Lysate at 40 ug, Primary: Anti-phospho-ROCK1 (Thr455+Ser456) (orb100140) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 158 kD, Observed band size: 158 kD.

Tissue/cell: human lung carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, 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-ROCK1 (Thr455/Ser456) Polyclonal Antibody, Unconjugated (orb100140) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.