Phospho-STAT3 (Tyr705) Rabbit Polyclonal Antibody

• Catalog Number: orb106147
• Category: Antibodies
• Description: Phospho-STAT3 (Tyr705) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Guinea pig, Porcine, Rabbit, Sheep
• Reactivity: Gallus, Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human STAT3 around the phosphorylation site of Tyr705 AP(p-Y)LK
• 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: 85 kDa
• Target: STAT3
• UniProt ID: P40763
• 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: STAT3 (phospho Y705); p-STAT3 (phospho Y705); Phos
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: A431. Primary Antibody (green line): Rabbit Anti-phospho-STAT3 (Tyr705) antibody (orb106147), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, 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°C. 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.

Paraformaldehyde-fixed, paraffin embedded (mouse heart), 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-STAT3 (Tyr705)) Polyclonal Antibody, Unconjugated (orb106147) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse pancreas), 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-STAT3 (Tyr705)) Polyclonal Antibody, Unconjugated (orb106147) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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-STAT3 (Tyr705)) Polyclonal Antibody, Unconjugated (orb106147) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat pancreas), 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-STAT3 (Tyr705)) Polyclonal Antibody, Unconjugated (orb106147) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Transplantation tumor), 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 (Phosphorylated signal Transductor and Activator of Transcription 3, p-STAT3) Polyclonal Antibody, Unconjugated (orb106147) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 minutes and DAB staining.

Sample: Heart (Mouse) Lysate at 40 ug, Heart (Rat) Lysate at 40 ug, Primary: Anti-p-STAT3 (orb106147) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 85 kD, Observed band size: 85 kD.

Sample: Lane 1: Heart (Mouse) Lysate at 40 ug, Lane 2: Kidney (Mouse) Lysate at 40 ug, Lane 3: Aorta (Rat) Lysate at 40 ug, Primary: Anti-phospho-STAT3 (Tyr705) (orb106147) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 92/84 kD, Observed band size: 84 kD.

The blue histogram is unstained cells (mouse kidney). The Orange histogram is cells stained with Rabbit IgG/FITC (orb525556)isotype control antibody. The green histogram is cells stained with Rabbit Anti-phospho-STAT3 (Tyr705)/FITC Conjugated antibody (orb16213). Concentration: 2 µg/10^6 cells or 5 µg/10^6 cells.

Human Melanoma cell line, EST02, was stained with Rabbit Anti-phospho-STAT3 (Tyr705) Polyclonal Antibody, FITC Conjugated (orb16213) for 30 minutes, on ice.

Tissue/Cell: human gastric cancer, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-phospho-STAT3 Polyclonal Antibody, Unconjugated (orb106147) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.