Phospho-P53 (Thr55) Rabbit Polyclonal Antibody

• Catalog Number: orb6604
• Category: Antibodies
• Description: Phospho-P53 (Thr55) Rabbit Polyclonal Antibody
• Target: TP53
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Purification: Affinity purified by Protein A
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human P53 around the phosphorylation site of Thr55 WF(p-T)ED
• UniProt ID: P04637
• RRID: AB_10938793
• MW: 43 kDa
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P
• Dilution range: IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Antibody Type: Primary Antibody
• 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.
• Alternative names: p53(phospho-Thr55); P53(phospho T55); P53(phospho
• Research Area: Cancer Research, Tumor Suppressors, p53 Signaling
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

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 (Thr55)) polyclonal Antibody, Unconjugated (orb6604) 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.

Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody (green line): Rabbit Anti-phospho-P53 (Thr55) antibody (orb6604), 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.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-phospho-P53 (Thr55) antibody (orb6604), 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 cervical 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 (p-P53 (Thr55)) Polyclonal Antibody, Unconjugated (orb6604) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human colon 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 (phospho-P53 (Thr55)) Polyclonal Antibody, Unconjugated (orb6604) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 minutes and DAB staining.