bsm-52210R [Primary Antibody]
Rabbit  Anti-phospho-STAT3 (Ser727)   Antibody
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Host: Rabbit

Target Protein: phospho-STAT3 (Ser727)

IR: Immunogen Range:PM(p-S)PR

Clonality:

Isotype: IgG

Entrez Gene: 6774

Swiss Prot: P40763

Source: KLH conjugated Synthesised phosphopeptide derived from human STAT3 around the phosphorylation site of Ser727:PM(p-S)PR 

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: STATs (signal transducers and activators of transcription) were originally discovered as two proteins (STAT1 and STAT2) which were involved in interferon alpha (IFN alpha) and IFN gamma signal transduction. Since then, several additional STAT proteins have been identified (STAT3, 4, 5a, 5b, and 6). STATs undergo tyrosine phosphorylation in response to growth factor or cytokine signaling. This phosphorylation results in dimerization and translocation of STAT proteins to the nucleus. In some cases this process is mediated by JAK Kinases (Janus Kinases 1, 2, and 3). For maximum activation of these proteins, phosphorylation at specific tyrosine and serine residues may be required in STAT1 alpha, 3, 4, and 5. Specific functions of the various members of the STAT family are poorly understood. STAT3 has been shown to be activated by IFN alpha but not IFN beta. The transcription factors associated with STAT3 are cJun and cyclic AMP responsive enhancer binding protein (CREB). Deletion of the STAT3 gene in knock out mice was lethal at the early embryonic stage.

Size: 100ul

Concentration: 1mg/ml

Applications: WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-200,Flow-Cyt=2ug/Test,ICC/IF=1:50-100,IF=1:100-500

Cross Reactive Species: Human,Mouse,Rat

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

VALIDATION IMAGES
Sample: Lane 1: Hela (Human) Cell Lysate at 30 ug Lane 2: A549 (Human) Cell Lysate at 30 ug Primary: Anti- phospho-STAT3 (Ser727) (bsm-52210R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 92/84 kD Observed band size: 92/84 kD
Sample: Heart (Rat) Lysate at 40 ug Cerebrum (Rat) Lysate at 40 ug Primary: Anti- phospho-STAT3 (Ser727) (bsm-52210R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 98 kD Observed band size: 94 kD
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boilingin 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boilingin 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); 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; Incubation with (phospho-STAT3 (Ser727) ) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human lung carcinoma); 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human lung); 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat lung); Antigen retrieval by boiling inin 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse lung); 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat pancreas); 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); 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; Incubation with (phospho-STAT3 (Ser727)) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-STAT3 (Ser727)) monoclonal Antibody, Unconjugated (bs-52210R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Blank control: A431. Primary Antibody (green line): Rabbit Anti-phospho-STAT3 (Ser727) antibody (bsm-52210R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-AF488 Dilution: 1μg /test. Protocol The cells were fixed with 4% PFA (10min 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.
 
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