TP53 Rabbit Polyclonal Antibody

• Catalog Number: orb573633
• Category: Antibodies
• Description: Rabbit polyclonal antibody to p53
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ChIP, ELISA, WB
• Predicted Reactivity: Human
• Reactivity: Human
• Immunogen: The immunogen is a synthetic peptide directed towards the N terminal region of human TP53
• Concentration: 0.5 mg/ml
• Form/Appearance: Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
• Conjugation: Unconjugated
• MW: 44 kDa
• Target: TP53
• UniProt ID: P04637
• Protein Sequence: Synthetic peptide located within the following region: EEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIE
• NCBI: NP_000537
• 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: Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
• Alternative names: P53, BCC7, LFS1, BMFS5, TRP53
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

25 ug of the indicated Human whole cell extracts was loaded onto a 12% SDS-PAGE gel. 1 ug/ml of the antibody was used in this experiment. TP53 is expressed as several different isoforms (> 10) ranging in size from 24 kDa to ~50 kDa in a cell type specific manner. This immunogen for this antibody is present in several isoforms. 1-3 ug/ml is recommended for this antibody.

DU145 cells were lysed in IP lysis buffer (20mM HEPES, 1% Triton X-100, 150mM NaCl, 1mMEDTA, 1mM EGTA, 100mM NaF, 10mM Na4P2O7, 1mM Na3VO4, 0.2mM PMSF). Amount of protein per well: 30 ug, Primary antibody conditions: 1:2000 in 5% milk/TBST buffer, overnight at 4°C.Secondary antibody conditions: 1:5000 in 5% milk/TBST buffer, 1 hour at room temperature. TP53 is strongly supported by BioGPS gene expression data to be expressed in Human DU145 cells.

Sample type: HeLa cells Antibody titration: 1 to 1000, Gel: 4-15% gradient, Secondary: Goat anti-rabbit HRP, Secondary dilution: 1 to 5000. TP53 is supported by BioGPS gene expression data to be expressed in HeLa.

U20S (p53+) cells were treated with 0.5 uM Doxorubicin for 14 hrs to induce DNA damage and hence activate p53. In parallel, PLKO cells (U2OS cells with stable shRNA-mediated knockdown of p53) were treated similarly and were used as negative control. Thedata for p21 promoter were normalised to actin (control for non-specific binding of DNA to the antibodies).

WB Suggested Anti-TP53 Antibody Titration: 0.2-1 ug/ml, ELISA Titer: 1:1562500, Positive Control: 293T. TP53 is strongly supported by BioGPS gene expression data to be expressed in HEK293T.