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Western blot analysis of p53 was performed by loading 40 µg of Nuclear protein extract from BEAS-2B in sample buffer and 5µL Spectra Multicolor Broad Range Protein Ladder(Product # 26634) onto a 10% polyacrylamide gel. Proteins were transferred to PVDF membrane (Product # 88518) at 65V for 90 min on ice. Membrane was blocked in 5% Milk in PBST for 1 hour at room temperature. p53 was detected at approximately 70 kDa using p53 monoclonal antibody (Product # 13-4100) at a dilution of 1:1500 in 0.5% Milk in PBST overnight at 4°C. Followed by goat anti-mouse alkaline phosphatase secondary antibody at a dilution of 1:20,000. Chemiluminescent detection was performed using ECF substrate. Data courtesy of Dr. Fondufe-Mittendorf's lab..
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant p53 peptide.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:1500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
13-4100 was successfully used to detect p53 in WB, IHC and IP applications.
Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. In cooperation with mitochondrial PPIF is involved in activating oxidative stress-induced necrosis; the function is largely independent of transcription. Induces the transcription of long intergenic non-coding RNA p21 (lincRNA-p21) and lincRNA- Mkln1. LincRNA-p21 participates in TP53-dependent transcriptional repression leading to apoptosis and seem to have to effect on cell-cycle regulation. Implicated in Notch signaling cross-over. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis.
IP-MS enrichment of TP53 (LFQ intensity): TP53 was enriched 3020-fold from BT549 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Product # 90409) and TP53 antibody (Product # 13-4100). STRING database was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
UV-induced ablation of the epidermal basal layer including p53-mutant clones resets UV carcinogenesis showing squamous cell carcinomas to originate from interfollicular epidermis.
13-4100 was used in immunohistochemistry to investigate the origin of UV-induced squamous cell carcinomas.
|Rebel HG,Bodmann CA,van de Glind GC,de Gruijl FR||Carcinogenesis (33:714)||2012|
Ionizing radiation-induced apoptosis via separate Pms2- and p53-dependent pathways.
13-4100 was used in western blot to test if p53 and MMR mediate X-ray cytotoxicity via the same pathway.
|Zeng M,Narayanan L,Xu XS,Prolla TA,Liskay RM,Glazer PM||Cancer research (60:4889)||2000|
Antigen NY-CO-13; Cellular tumor antigen p53; mutant p53; mutant tumor protein 53; p53 cellular tumor antigen; p53 tumor suppressor; Phosphoprotein p53; TP53; transformation related protein 53; transformation-related protein 53; tumor protein 53; tumor protein p53 (Li-Fraumeni syndrome); Tumor suppressor p53; tumor supressor p53
bbl; BCC7; bfy; bhy; LFS1; p44; P53; TP53; TRP53