Anti-Phospho-P53 (Ser33) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-P53 (Ser33). Anti-Phospho-P53 (Ser33) Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat

1. Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-phospho-P53 (Ser33) antibody (TMAB-11081), Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg /test.
2. Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); 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-P53 (Ser33)) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. 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-P53 (Ser33)) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (mouse bladder); 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-P53 (Ser33)) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
5. Sample:
Urinary bladder (Mouse) Lysate at 40 μg
Skin (Mouse) Lysate at 40 μg
Primary: Anti-phospho-P53 (TMAB-11081) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kD
Observed band size: 53 kD
6. Blank control: A549.
Primary Antibody (green line): Rabbit Anti-phospho-P53 (Ser33) antibody (TMAB-11081)
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℃. 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 20,000 events was performed.
7. A549 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-P53 (Ser33)) polyclonal Antibody, Unconjugated (TMAB-11081) 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 nuclei.
8. Paraformaldehyde-fixed, paraffin embedded Mouse Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
13. Paraformaldehyde-fixed, paraffin embedded Human Ovarian Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
14. Paraformaldehyde-fixed, paraffin embedded Human Skin Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
15. Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
16. Paraformaldehyde-fixed, paraffin embedded Human Esophagus Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
17. Paraformaldehyde-fixed, paraffin embedded Human Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-P53 (Ser33) Polyclonal Antibody, Unconjugated (TMAB-11081) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:50-200; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human P53 around the phosphorylation site of Ser33

Human

TP53

Cellular tumor antigen p53

Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. Implicated in Notch signaling cross-over. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis.