Antibody Type: Rabbit Polyclonal

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

Reactivity: Human,Mouse,Rat (predicted:Dog,Pig,Horse,Rabbit,Sheep)

IgG

Human,Mouse,Rat (predicted:Dog,Pig,Horse,Rabbit,Sheep)

1. Tissue/cell: rat brain tissue; 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-P53 (wt-p53) Polyclonal Antibody, Unconjugated (TMAB-01320) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Blank control (blue line): Hela (blue).
Primary Antibody (green line): Rabbit Anti-P53 (wt-p53) antibody (TMAB-01320),
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.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. Sample:
Liver (Mouse) Lysate at 40 μg
Liver (Rat) Lysate at 40 μg
Primary: Anti-P53 (wt-p53) (TMAB-01320) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
4. Tissue/cell: 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 (P53 (wt-p53)) polyclonal Antibody, Unconjugated (TMAB-01320) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
5. Tissue/cell: 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 (P53 (wt-p53)) polyclonal Antibody, Unconjugated (TMAB-01320) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
6. Sample:
MCF-7 (Human) Cell Lysate at 40 μg
DU145 (Human) Cell Lysate at 40 μg
Primary: Anti-P53 (wt-p53) (TMAB-01320) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa

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

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

Polyclonal

KLH conjugated synthetic peptide: human P53

Human

7157

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