Antibody Type: Rabbit Polyclonal

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

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

IgG

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

1. Paraformaldehyde-fixed, paraffin embedded (Rat bladder); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Transformation related protein 53; P53) Polyclonal Antibody, Unconjugated (TMAB-01321) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 min and DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (Rat bladder); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Transformation related protein 53; P53) Polyclonal Antibody, Unconjugated (TMAB-01321) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 min and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (Rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Transformation related protein 53; P53) Polyclonal Antibody, Unconjugated (TMAB-01321) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 min and DAB staining.
4. Sample: 293T Cell (Human) Lysate at 40 μg
Primary: Anti-p53 (FL-393) (TMAB-01321) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 60 kDa
5. Sample: MCF-7 Cell (Human) Lysate at 40 μg
Primary: Anti-p53 (FL-393) (TMAB-01321) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 60 kDa
6. Paraformaldehyde-fixed, paraffin embedded (mouse intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (p53 (FL-393)) Polyclonal Antibody, Unconjugated (TMAB-01321) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded (mouse lung); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (p53 (FL-393)) Polyclonal Antibody, Unconjugated (TMAB-01321) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded (rat skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (p53 (FL-393)) Polyclonal Antibody, Unconjugated (TMAB-01321) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded (mouse skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (p53 (FL-393)) Polyclonal Antibody, Unconjugated (TMAB-01321) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
10. Blank control: A549. Primary Antibody (green line): Rabbit Anti-p53 (FL-393) antibody (TMAB-01321)
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 room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
11. 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 (p53 (FL-393)) polyclonal Antibody, Unconjugated (TMAB-01321) 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.

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

WB: 1:1000-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1ug/Test

Polyclonal

Full length P53 protein of human origin

Human

7157

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