P53 protein (wt-p53) Rabbit Polyclonal Antibody

• Catalog Number: orb11210
• Category: Antibodies
• Description: P53 protein (wt-p53) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Equine, Porcine, Rabbit, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human P53 (251-310/393aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 43 kDa
• Target: TP53
• UniProt ID: P04637
• RRID: AB_10750207
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Widespread p53; Wtp53; Antigen NY-CO-13; Cellular
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Wittmann, Franziska et al. To Be Or Not to Be: the "Smoker's Paradox" - An in-Vitro Study Cell Physiol Biochem, 48, 1638-1651 (2018)

PubMed: 30077999

Blank control (blue line): HeLa (blue). Primary Antibody (green line): Rabbit Anti-P53 protein (wt-p53) antibody (orb11210), 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. Acquisition of 20000 events was performed.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-P53 protein (wt-p53) antibody (orb11210), 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. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Human breast cancer), 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 (P53 protein (wt-p53)) Polyclonal Antibody, Unconjugated (orb11210) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse esophageal), 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 (P53 protein (wt-p53)) Polyclonal Antibody, Unconjugated (orb11210) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (P53 protein (wt-p53)) Polyclonal Antibody, Unconjugated (orb11210) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat esophageal), 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 (P53 protein (wt-p53)) Polyclonal Antibody, Unconjugated (orb11210) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Sample: Brain (Rat) lysate at 30 ug, Colon carcinoma (Human) lysate at30 ug, Primary: Anti-wt-p53 (orb11210) at 1:200 dilution, Secondary: HRP conjugated Goat-Anti-Rabbit IgG at 1:3000 dilution, Predicted band size: 43kD, Observed band size: 53kD.

Sample: HepG2 Cell Lysate at 40 ug, Mcf-7 Cell Lysate at 40 ug, DU145 Cell Lysate at 40 ug, Primary: Anti-P53 protein (wt-p53) (orb11210) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 43 kD, Observed band size: 50 kD.

Sample: Liver (Mouse) Lysate at 40 ug, Primary: Anti-P53 protein (wt-p53) (orb11210) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 43 kD, Observed band size: 50 kD.

Tissue/Cell: 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 protein (wt-p53)) polyclonal Antibody, Unconjugated (orb11210) 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 nuclei.

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 protein (wt-p53)) polyclonal Antibody, Unconjugated (orb11210) 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 nuclei.

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 protein (wt-p53)) polyclonal Antibody, Unconjugated (orb11210) 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 nuclei.