Phospho-TP53 (S392) Recombinant Monoclonal Antibody
Description
Definition and Purpose of Phospho-TP53 (S392) Recombinant Monoclonal Antibody
The Phospho-TP53 (S392) Recombinant Monoclonal Antibody is a highly specific immunological tool designed to detect phosphorylation at serine 392 (S392) of the tumor suppressor protein p53 (encoded by the TP53 gene). This post-translational modification is critical for regulating p53’s stability, DNA-binding activity, and transcriptional output, particularly in response to cellular stress or DNA damage . The antibody is engineered using recombinant technology to ensure high specificity and reproducibility, targeting a phosphorylated peptide epitope near S392 .
Detection of Phosphorylated TP53 in Cellular Stress Models
The antibody is widely used to study TP53 activation under genotoxic stress. For example:
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Camptothecin treatment: MCF-7 breast cancer cells treated with 1 µM camptothecin (CPT) show a robust 53 kDa band corresponding to phosphorylated TP53 (S392) in Western blot .
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Actinomycin D exposure: MCF-7 cells treated with 5 µg/ml actinomycin for 3–18 hours exhibit time-dependent phosphorylation at S392, detected via WB .
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Alkaline Phosphatase Treatment: Membranes treated with alkaline phosphatase lose the 53 kDa band, confirming phospho-specificity .
Comparative Performance Across Cell Lines
Immunohistochemistry and Localization
The antibody detects cytoplasmic and nuclear TP53 phosphorylation in paraffin-embedded tissues:
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Prostate adenocarcinoma: Strong nuclear staining with clone EP155Y .
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Ovarian/Gastric carcinomas: Cytoplasmic and nuclear localization observed with clone EP1889Y .
Antibody Validation and Optimization
| Parameter | Optimal Conditions |
|---|---|
| Western Blot Dilution | 1:500–1:2000 (varies by cell lysate concentration) |
| IHC Dilution | 1:50–1:200 (antigen retrieval with citrate buffer pH 6 recommended) |
| Blocking Buffer | 5% non-fat dry milk (NFDM) or BSA in TBST |
| Secondary Antibody | HRP-conjugated anti-rabbit IgG (1:10,000–1:20,000) |
| Critical Controls | Lambda-phosphatase-treated lysates (negative control) |
Clinical and Biological Relevance
Phosphorylation at S392 is implicated in:
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Tumor Formation: Hyperphosphorylation correlates with human tumors, as TP53 mutations disrupt normal regulatory pathways .
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Therapeutic Targeting: The antibody aids in identifying TP53 phosphorylation dynamics for drug development (e.g., stabilizing agents or kinase inhibitors) .
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Stress Response: S392 phosphorylation modulates TP53’s interaction with coactivators (e.g., p300) and repressors (e.g., MDM2), influencing cell cycle arrest or apoptosis .
Product Specs
The phospho-TP53 (S392) recombinant monoclonal antibody is produced through a meticulous process involving the isolation of antibody-coding genes from rabbits previously exposed to a synthesized peptide derived from the human TP53 protein phosphorylated at S392. These genes are then carefully integrated into specialized expression vectors. Subsequently, these vectors are introduced into host suspension cells, which are cultivated to promote the production and secretion of the antibody. Following this cultivation phase, the phospho-TP53 (S392) recombinant monoclonal antibody undergoes a stringent purification process utilizing affinity chromatography techniques, effectively separating the antibody from the cell culture supernatant. Finally, its functionality is rigorously assessed through a comprehensive battery of tests, including ELISA and WB, confirming its ability to specifically interact with the human TP53 protein phosphorylated at S392.
Target Background
TP53 acts as a tumor suppressor in numerous tumor types, inducing growth arrest or apoptosis based on the specific physiological conditions and cell type. It plays a crucial role in cell cycle regulation as a trans-activator that negatively regulates cell division by controlling genes essential for this process. Notably, one of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction is likely mediated through stimulation of BAX and FAS antigen expression or by repressing Bcl-2 expression. Its pro-apoptotic activity is activated upon interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2. However, this activity is inhibited when the interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 is displaced by PPP1R13L/iASPP. In collaboration with mitochondrial PPIF, TP53 participates in activating oxidative stress-induced necrosis, largely independent of transcription. It induces the transcription of long intergenic non-coding RNA p21 (lincRNA-p21) and lincRNA-Mkln1. LincRNA-p21 is involved in TP53-dependent transcriptional repression leading to apoptosis and appears to influence cell-cycle regulation. TP53 is also implicated in Notch signaling cross-over. When associated with the CAK complex in response to DNA damage, TP53 prevents CDK7 kinase activity, thereby halting cell cycle progression. Isoform 2 enhances the transactivation activity of isoform 1 from some, but not all, TP53-inducible promoters. Conversely, isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis. TP53 regulates the circadian clock by repressing CLOCK-ARNTL/BMAL1-mediated transcriptional activation of PER2.
- This review summarizes the diverse functions of p53 in adipocyte development and adipose tissue homeostasis. It also explores the manipulation of p53 levels in adipose tissue depots and their impact on systemic energy metabolism in the context of insulin resistance and obesity. PMID: 30181511
- This study reveals a USP15-dependent lysosomal pathway controlling p53-R175H turnover in ovarian cancer cells. PMID: 29593334
- The findings indicate that the underlying mechanisms by which etoposide and ellipticine regulate CYP1A1 expression differ and may not solely rely on p53 activation. PMID: 29471073
- This study investigated the association of tumor protein p53 and drug metabolizing enzyme polymorphisms with clinical outcomes in patients with advanced nonsmall cell lung cancer. PMID: 28425245
- POH1 knockdown induced cell apoptosis through increased expression of p53 and Bim. PMID: 29573636
- This study reveals a previously unrecognized effect of chronic high-fat diet on beta-cells, where persistent oxidative stress results in p53 activation and subsequent inhibition of mRNA translation. PMID: 28630491
- Diffuse large B cell lymphoma lacking CD19 or PAX5 expression were more likely to have mutant TP53. PMID: 28484276
- This research demonstrates that proliferation potential-related protein promotes esophageal cancer cell proliferation and migration, and suppresses apoptosis by mediating the expression of p53 and IL-17. PMID: 30223275
- HIV-1 infection and subsequent HIV-1 reverse transcription are inhibited in HCT116 p53(+/+) cells compared to HCT116 p53(-/-) cells. Tumor suppressor gene p53 expression is upregulated in non-cycling cells. The restrictions of HIV by p53 are associated with the suppression of ribonucleotide reductase R2 subunit expression and phosphorylation of SAMHD1 protein. PMID: 29587790
- The study highlights that MDM2 and MDMX are targetable vulnerabilities within TP53-wild-type T-cell lymphomas. PMID: 29789628
- A significant increase in the expression of p53 and Bax was observed in cells treated with alpha-spinasterol, while cdk4/6 were significantly down-regulated upon exposure to alpha-spinasterol. PMID: 29143969
- A significant correlation was found between telomere dysfunction indices, p53, oxidative stress indices, and malignant stages of GI cancer patients. PMID: 29730783
- PGEA-AN modulates the P53 system, leading to the death of neuroblastoma cells without affecting the renal system in vivo. This suggests its potential as a future prospect for developing an anticancer moiety against neuroblastoma. PMID: 29644528
- These data indicate that activation of autophagy reduces the expression of STMN1 and p53, and the migration and invasion of cancer cells. This contributes to the anti-cancer effects of Halofuginone. These findings may provide new insight into breast cancer prevention and therapy. PMID: 29231257
- miR-150 suppresses cigarette smoke-induced lung inflammation and airway epithelial cell apoptosis, causally linked to repression of p53 expression and NF-kappaB activity. PMID: 29205062
- Tumors harboring TP53 mutations, which can impair epithelial function, exhibit a unique bacterial consortium that is relatively more abundant in smoking-associated tumors. PMID: 30143034
- This review explores the intricate crosstalk among p53, lipid metabolism, insulin resistance, inflammation, and oxidative stress, highlighting their roles in non-alcoholic fatty liver disease. PMID: 30473026
- Ubiquitin-conjugating enzyme E2S (UBE2S) enhances the ubiquitination of p53 protein, facilitating its degradation in hepatocellular carcinoma (HCC) cells. PMID: 29928880
- p53 knockout compensates for osteopenia in murine Mysm1 deficiency. PMID: 29203593
- SIRT1 plays a pivotal protective role in regulating ADSCs aging and apoptosis induced by H2O2. PMID: 29803744
- 133p53 promotes tumor invasion via IL-6 through activation of the JAK-STAT and RhoA-ROCK pathways. PMID: 29343721
- Mutant TP53 G245C and R273H can lead to more aggressive phenotypes and enhance cancer cell malignancy. PMID: 30126368
- PD-L1, Ki-67, and p53 staining individually hold significant prognostic value for patients with stage II and III colorectal cancer. PMID: 28782638
- This study of patients with ccRCC, using pooled analysis and multivariable modeling, demonstrates that three recurrently mutated genes, BAP1, SETD2, and TP53, have statistically significant associations with poor clinical outcomes. Importantly, mutations of TP53 and SETD2 were associated with decreased CSS and RFS, respectively. PMID: 28753773
- This study reveals that the Wnt/beta-catenin signaling pathway and its primary downstream target, c-Myc, increase miR552 levels. miR552 directly targets the p53 tumor suppressor. Thus, miR552 may serve as a crucial link between functional loss of APC, leading to abnormal Wnt signals, and the absence of p53 protein in colorectal cancer. PMID: 30066856
- High levels of glucose induce endothelial dysfunction through TAF1-mediated p53 Thr55 phosphorylation and subsequent GPX1 inactivation. PMID: 28673515
- While tumor protein p53 (p53) does not directly control the luminal fate, its loss facilitates the acquisition of mammary stem cell (MaSC)-like properties by luminal cells. This predisposes them to developing mammary tumors with loss of luminal identity. PMID: 28194015
- Fifty-two percent of patients diagnosed with glioma/glioblastoma exhibited a positive TP53 mutation. PMID: 29454261
- The expression of Ser216pCdc25C was also elevated in the combined group, indicating that irinotecan likely radiosensitized the p53-mutant HT29 and SW620 cells through the ATM/Chk/Cdc25C/Cdc2 pathway. PMID: 30085332
- In the former, p53 binds to the CDH1 (encoding E-cadherin) locus to antagonize EZH2-mediated H3K27 trimethylation (H3K27me3), maintaining high levels of acetylation of H3K27 (H3K27ac). PMID: 29371630
- Among the hits, miR-596 was identified as a regulator of p53. Overexpression of miR-596 significantly increased p53 at the protein level, inducing apoptosis. PMID: 28732184
- Apoptosis pathways are impaired in fibroblasts from patients with SSc, leading to chronic fibrosis. However, the PUMA/p53 pathway may not be involved in the dysfunction of apoptosis mechanisms in fibroblasts of patients with SSc. PMID: 28905491
- Low TP53 expression is associated with drug resistance in colorectal cancer. PMID: 30106452
- The activation of p38 in response to low doses of ultraviolet radiation is hypothesized to be protective for p53-inactive cells. Therefore, MCPIP1 may favor the survival of p53-defective HaCaT cells by sustaining the activation of p38. PMID: 29103983
- TP53 missense mutations are associated with castration-resistant prostate cancer. PMID: 29302046
- P53 degradation is mediated by COP1 in breast cancer. PMID: 29516369
- Combined inactivation of the XRCC4 non-homologous end-joining (NHEJ) DNA repair gene and p53 efficiently induces brain tumors with hallmark characteristics of human glioblastoma. PMID: 28094268
- This study establishes a direct link between Y14 and p53 expression, suggesting a function for Y14 in DNA damage signaling. PMID: 28361991
- TP53 Mutation is associated with Mouth Neoplasms. PMID: 30049200
- Cryo-Electron Microscopy studies on p53-bound RNA Polymerase II (Pol II) reveal that p53 structurally regulates Pol II, influencing its DNA binding and elongation. This provides new insights into p53-mediated transcriptional regulation. PMID: 28795863
- Increased nuclear p53 phosphorylation and PGC-1alpha protein content immediately following SIE but not CE suggests these may represent significant early molecular events in the exercise-induced response to exercise. PMID: 28281651
- The E6/E7-p53-POU2F1-CTHRC1 axis promotes cervical cancer cell invasion and metastasis. PMID: 28303973
- Accumulated mutant-p53 protein suppresses the expression of SLC7A11, a component of the cystine/glutamate antiporter, system xC(-), by binding to the master antioxidant transcription factor NRF2. PMID: 28348409
- Consistently, forced expression of p53 significantly stimulated ACER2 transcription. Notably, p53-mediated autophagy and apoptosis were markedly enhanced by ACER2. Depletion of the essential autophagy gene ATG5 revealed that ACER2-induced autophagy facilitates its effect on apoptosis. PMID: 28294157
- These results indicate that LGASC of the breast is a low-grade triple-negative breast cancer that harbors a basal-like phenotype with no androgen receptor expression and exhibits a high rate of PIK3CA mutations but no TP53 mutations. PMID: 29537649
- This study demonstrates an inhibitory effect of wild-type P53 gene transfer on graft coronary artery disease in a rat model. PMID: 29425775
- Our results suggest that TP53 c.215G>C, p. (Arg72Pro) polymorphism may be considered a genetic marker for predisposition to breast cancer in the Moroccan population. PMID: 29949804
- Higher levels of the p53 isoform, p53beta, predict better prognosis in patients with renal cell carcinoma through enhancing apoptosis in tumors. PMID: 29346503
- TP53 mutations are associated with colorectal liver metastases. PMID: 29937183
- High expression of TP53 is associated with oral epithelial dysplasia and oral squamous cell carcinoma. PMID: 29893337
