Antibody Type: Recombinant Rabbit Monoclonal

Application: WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

Reactivity: Human,Rat,Mouse

IgG

8J881

Human,Rat,Mouse

1. Sample:
Heart (Rat) Lysate at 40 μg
Cerebrum (Rat) Lysate at 40 μg
Primary: Anti-phospho-STAT3 (Ser727) (TMAB-01507) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 98 kDa
Observed band size: 94 kDa
2. Blank control: A431. Primary Antibody (green line): Rabbit Anti-phospho-STAT3 (Ser727) antibody (TMAB-01507)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
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 at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. Sample:
Lane 1: Hela (Human) Cell Lysate at 30 μg
Lane 2: A549 (Human) Cell Lysate at 30 μg
Primary:
Anti-phospho-STAT3 (Ser727) (TMAB-01507) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 92/84 kDa
Observed band size: 92/84 kDa
4. A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (phospho-STAT3 (Ser727)) monoclonal Antibody, Unconjugated 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
5. Paraformaldehyde-fixed, paraffin embedded Mouse Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded Rat Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
9. Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
10. Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
13. Paraformaldehyde-fixed, paraffin embedded Mouse Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
14. Paraformaldehyde-fixed, paraffin embedded Mouse Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
15. Paraformaldehyde-fixed, paraffin embedded Rat Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
16. Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
17. Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
18. Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
19. Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (TMAB-01507) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

WB: 1:500-2000; IHC-P: 1:50-200; IHC-Fr: 1:50-200; ICC/IF: 1:50-200; IF: 1:50-200; FCM: 1:20-50

Monoclonal

A synthesized peptide: human STAT3 around the phosphorylation site of S727

Human

6774

STAT family,Transcription factors/regulators,Transcription factors,Surface molecules,STATs,Other factors,SARS Coronavirus,STATs,Intracellular