Acetyl-PTEN (K402) Antibody
Description
Fundamental Characteristics of Acetyl-PTEN (K402) Antibody
Acetyl-PTEN (K402) antibody is a polyclonal antibody derived from rabbit immunized with synthesized peptides containing the acetylated lysine 402 of human PTEN protein. This antibody specifically recognizes PTEN when acetylated at the K402 position, making it an invaluable tool for studying post-translational modifications of the PTEN tumor suppressor. The antibody demonstrates cross-reactivity with human, mouse, and rat samples, making it versatile for comparative studies across species .
The immunogen used for antibody generation consists of synthesized acetyl-peptides derived specifically from the human PTEN protein around the acetylation site of K402. The antibody recognizes the modification sequence "QITkV," where the lowercase "k" represents the acetylated lysine residue . This specific recognition pattern ensures high selectivity for the acetylated form of PTEN, allowing researchers to distinguish between acetylated and non-acetylated states of the protein.
Production and Purification Methods
The antibody is produced through immunization of rabbits with the synthetic acetylated peptide, followed by affinity purification. The purification process typically involves affinity chromatography using the epitope-specific immunogen, which substantially increases the specificity of the antibody by removing non-specific antibodies from the rabbit antiserum . This rigorous purification method ensures that the resulting antibody preparation maintains high specificity for the acetylated K402 site of PTEN.
PTEN Structure and the Significance of K402 Acetylation
PTEN (Phosphatase and Tensin homolog) is a critical tumor suppressor protein that functions primarily as a lipid phosphatase, specifically dephosphorylating the 3-position of phosphatidylinositols. This activity directly counteracts the phosphatidylinositol 3-kinase (PI3K) pathway, which is activated by numerous growth-stimulating signals . Understanding the structure of PTEN provides context for the significance of K402 acetylation.
Regulatory Role of K402 Acetylation
K402 acetylation represents one of several post-translational modifications that regulate PTEN function. Unlike acetylation at K125 and K128, which directly affects the catalytic activity by modifying residues at the catalytic center, K402 acetylation primarily influences PTEN's protein-protein interactions . The K402 residue is located in the PDZ-binding domain, and its acetylation modulates how PTEN interacts with other PDZ domain-containing proteins .
Research has identified CBP (CREB-binding protein) as the primary acetyltransferase responsible for K402 acetylation, while SIRT1 functions as the main deacetylase that removes the acetyl group from this position . This dynamic acetylation-deacetylation process provides a regulatory mechanism for controlling PTEN's interactions with its binding partners without directly affecting its phosphatase activity .
Applications of Acetyl-PTEN (K402) Antibody in Research
The Acetyl-PTEN (K402) antibody serves as a valuable tool in various research applications focusing on post-translational modifications of PTEN and their functional implications. The primary applications include:
Western Blot Analysis
Western blotting represents the most common application for Acetyl-PTEN (K402) antibody, allowing researchers to detect and quantify acetylated PTEN in cell and tissue lysates. The antibody effectively recognizes the acetylated form of PTEN at approximately 47 kDa on immunoblots . This application enables investigators to monitor changes in PTEN acetylation status under different experimental conditions, such as drug treatments or genetic manipulations.
For optimal results in Western blot applications, the recommended dilution range is typically 1:500 to 1:2000 . Secondary antibodies conjugated with horseradish peroxidase (HRP) are commonly used for detection, with enhanced chemiluminescence (ECL) systems providing sensitive visualization of the acetylated PTEN protein.
Enzyme-Linked Immunosorbent Assay (ELISA)
The Acetyl-PTEN (K402) antibody can be effectively utilized in ELISA systems for quantitative measurement of acetylated PTEN levels. For ELISA applications, the antibody is typically used at higher dilutions ranging from 1:2000 to 1:20000 . This application provides a high-throughput method for screening acetylated PTEN levels across multiple samples simultaneously.
Immunohistochemistry (IHC)
Immunohistochemical analysis of tissue sections represents another important application for the Acetyl-PTEN (K402) antibody. This technique allows researchers to visualize the distribution and localization of acetylated PTEN within tissues and cellular compartments. For IHC applications, the recommended dilution range is typically 1:50 to 1:300 . This application is particularly valuable for studying acetylated PTEN in tumor samples and comparing its expression patterns with normal tissues.
PTEN Acetylation in Cellular Signaling and Cancer Biology
The acetylation of PTEN at K402 plays a significant role in cellular signaling networks and has implications for cancer biology. Understanding these connections provides context for the importance of studying PTEN K402 acetylation.
Regulation of PTEN Localization and Protein Interactions
PTEN localization is crucial for its tumor suppressor function, as it must access its substrates at cellular membranes. While K402 acetylation does not directly affect PTEN's phosphatase activity, it significantly modulates protein-protein interactions through the PDZ-binding domain . These interactions influence PTEN's subcellular localization and stability, indirectly affecting its tumor suppressor function.
The PDZ-binding domain of PTEN interacts with several PDZ domain-containing proteins, and K402 acetylation can either enhance or inhibit these interactions depending on the specific binding partner. This provides a dynamic regulatory mechanism that responds to cellular signals and conditions .
PTEN Post-Translational Modifications in Cancer
PTEN is regulated by multiple post-translational modifications including phosphorylation, acetylation, SUMOylation, and ubiquitination. In addition to K402 acetylation, PTEN can be acetylated at K125, K128, and K163, each with distinct functional consequences . Understanding the interplay between these modifications is crucial for comprehending PTEN's role in cancer suppression.
Dysregulation of PTEN post-translational modifications, including acetylation, has been implicated in cancer development and progression. The ability to specifically detect K402 acetylation using the Acetyl-PTEN (K402) antibody provides researchers with a tool to investigate these regulatory mechanisms in various cancer types .
Experimental Considerations and Optimization
When working with Acetyl-PTEN (K402) antibody, several experimental considerations can help optimize results and ensure reliable data interpretation.
Sample Preparation Considerations
The detection of acetylated PTEN requires careful sample preparation to preserve the acetylation status of the protein. Inclusion of deacetylase inhibitors such as trichostatin A (TSA) or nicotinamide in lysis buffers is recommended to prevent deacetylation during sample processing. Additionally, phosphatase inhibitors should be included to maintain the phosphorylation state of PTEN, which may influence its acetylation and detection .
Controls and Validation
Proper experimental controls are essential when working with Acetyl-PTEN (K402) antibody. Positive controls may include samples treated with deacetylase inhibitors to increase PTEN acetylation levels, while negative controls might involve samples treated with acetyltransferase inhibitors or PTEN-null cell lines. Additionally, using PTEN K402R mutants (where lysine is replaced with arginine to prevent acetylation) can provide valuable control conditions for antibody specificity validation .
Recent Research Findings and Future Directions
Recent research has shed light on the functional significance of PTEN K402 acetylation in various cellular processes and disease states. These findings highlight the importance of tools like the Acetyl-PTEN (K402) antibody in advancing our understanding of PTEN regulation.
Functional Implications of K402 Acetylation
Studies have demonstrated that K402 acetylation affects PTEN's binding with PDZ domain-containing proteins without directly impacting its phosphatase activity . This suggests that acetylation at this site primarily modulates PTEN's protein interaction network rather than its enzymatic function. The dynamic regulation of these interactions by acetylation/deacetylation provides an additional layer of control over PTEN's cellular activities .
PTEN Acetylation in Cellular Compartmentalization
Research has revealed that post-translational modifications, including acetylation, influence PTEN's distribution between cellular compartments. PTEN can localize to the cytoplasm, nucleus, and membrane, with different modifications directing its trafficking between these compartments. While K402 acetylation primarily affects protein-protein interactions through the PDZ-binding domain, other acetylation sites like K163 more directly influence membrane translocation . The complex interplay between these modifications determines PTEN's subcellular localization and, consequently, its access to substrates and binding partners.
Product Specs
Target Background
- Nuclear phosphatase and tensin homologue on chromosome ten protein (PTEN) interacts with the splicing machinery, spliceosome, to regulate its assembly and pre-mRNA splicing. PMID: 29921876
- The expression of PTEN and miR-144 was inversely correlated in metastatic breast cancer cell lines. PMID: 30132256
- Disruption of PTEN protein isoform PTENbeta (PTENbeta) alters rDNA transcription and promotes ribosomal biogenesis. PMID: 28332494
- Functionally, Ataxin-3 overexpression promoted cell proliferation, and Ataxin-3 knockdown inhibited cell proliferation in testicular cancer cells. Furthermore, up-regulation of Ataxin-3 inhibited the expression of PTEN and activated the AKT/mTOR pathway. PMID: 29902454
- A certain degree of mitochondrial oxidative activity was observed, with some differences between PTEN-wild type SF767 cells and PTEN-deleted A172 and U87MG cells, which are characterized by a loss-of-function point mutation of PTEN. PMID: 29209894
- We demonstrated that expression of PTEN and miR-718 was significantly correlated in patients with gastric cancer. Low expression of PTEN and high levels of miR-718 were notably associated with a lower 5-year overall survival rate. Both PTEN and miR-718 were identified as prognostic factors of gastric cancer. PMID: 30131483
- The data suggest that diagnostic or therapeutic chest radiation may predispose patients with decreased stromal PTEN expression to secondary breast cancer, and that prophylactic EGFR inhibition may reduce this risk. PMID: 30018330
- Findings indicate that shikonin inhibits proliferation and promotes apoptosis in human endometrioid endometrial cancer (EEC) cells by modulating the miR-106b/PTEN/AKT/mTOR signaling pathway, suggesting that shikonin could act as a potential therapeutic agent in EEC treatment. PMID: 29449346
- SIRT6 inhibited proliferation, migration, and invasion of colon cancer cells by up-regulating PTEN expression and down-regulating AKT1 expression. PMID: 29957460
- Data show that phosphatase and tensin homolog (PTEN) interacts with death domain-associated protein (DAXX) and, in turn, PTEN directly regulates oncogene expression by modulating DAXX-histone H3.3 (H3.3) association on the chromatin. PMID: 28497778
- The study suggested that there may be a regulatory loop between miR21 and PTEN, and that miR21 inhibition affected the proliferative, invasive, and apoptotic abilities of oral squamous cell carcinoma (OSCC) cells. miR-21 expression was observed in 80.0% of OSCC tissues and in 30.0% of normal tissues. By contrast, PTEN expression exhibited an opposite trend in OSCC tissues (37.1%) and normal tissues (80.0%). PMID: 30132571
- MTSS1 is stabilized by the protein phosphatase activity of the tumor suppressor PTEN. Our data show that PTEN loss in PDAC cells results in both increased metastatic potential and decreased MTSS1 expression. Furthermore, we show that ectopic MTSS1 expression rescues this effect. PMID: 29175021
- Low PTEN mRNA expression was associated with down-regulation of a group of genes involved in immune responses and B-cell development. PMID: 29734016
- Results showed that MiR-374b was highly expressed, while PTEN was downregulated in the GIST tissues. The levels of miR-374b, PI3K, AKT, and PTEN were related to tumor diameter and pathological stage. Additionally, miR-374b increased the mRNA and protein levels of PI3K, Akt, MMP2, MMP9, P53, and cyclinD1, suggesting that miR-374b activates the PI3K/Akt signaling pathway in GIST-T1 cells. PMID: 29902839
- PTEN loss is associated with castration-resistant prostate cancer. PMID: 29302046
- Low PTEN expression is associated with thyroid cancer progression. PMID: 30015900
- We provide a review on current understandings of the regulation of PTEN by ncRNAs, which could contribute to the development of novel approaches to diseases with abnormal expression of PTEN. PMID: 30217221
- The IRIS-driven metastatic mechanism results from IRIS-dependent suppression of phosphatase and tensin homolog (PTEN) transcription, which in turn perturbs the PI3K/AKT/GSK-3beta pathway leading to prolyl hydroxylase-independent HIF-1alpha stabilization and activation in a normoxic environment. PMID: 30254159
- In this study, we used the Ion Personal Genome Machine (PGM) and Ion Torrent Ampliseq Cancer panel to sequence hotspot regions from PIK3CA, AKT, and PTEN genes to identify genetic mutations in 39 samples of TNBC subtype from Moroccan patients and to correlate the results with clinical-pathologic data. PMID: 30227836
- Data indicate a significant prognostic role for assessing transcriptional regulator ERG (ERG) and phosphatase and tensin homolog protein (PTEN) in men with prostate cancer. PMID: 30101374
- Low PTEN expression is associated with multiple myeloma. PMID: 30015974
- The loss of Sirt3 triggered fatal mitochondrial fission by suppressing the Akt/PTEN pathway. PMID: 30021354
- Results showed that SIX1 was overexpressed in osteosarcoma tissues, blood samples, and cell lines, whereas PTEN expression was reduced. PMID: 29807230
- miR23b3p and PTEN interfered with the viability and apoptosis of smooth muscle cells. PMID: 29845190
- PDCD4 and PTEN were the functional targets of miR-21. PMID: 30074182
- miR-205 functions as an oncogenic miRNA by directly binding to SMAD4 and PTEN, providing a novel target for the molecular treatment of ovarian cancer. PMID: 28145479
- Studies have indicated that in breast cancer, PTEN undergoes mutations. There is a functional and mechanistic link between the BMI-1 oncoprotein and tumor suppressor PTEN in the development and progression of breast cancer. [review] PMID: 30096458
- When considered together (43 cases), 1/25 cases (4%) with a PIK3CA mutation and/or low PTEN expression levels had a pathologic complete response (pCR) compared to 7/18 cases (39%) with wild-type PI3KCA and high PTEN expression levels (p = 0.006). PMID: 29110152
- Taken together, the authors presented here a novel cross-talk between miR-181a and PTEN, which was raised by hepatitis B virus X protein, and this shined a new line in hepatitis B virus-related hepato-carcinogenesis. PMID: 28053323
- Bioinformatics analysis demonstrated that the 3'UTR of PTEN mRNA was targeted by hsa-miR-142-5p, which regulates its expression, triggering cancer stem cell-like properties of cutaneous squamous cell carcinoma. PMID: 28857248
- PTEN lipid phosphatase inactivation abolished the MOB1-LATS1/2 interaction, decreased YAP phosphorylation, and finally promoted YAP nuclear translocation, which enhanced the synergistic effect of YAP-TEAD, thus inducing cell proliferation and migration. PMID: 30134988
- TERT could induce thyroid carcinoma cell proliferation mainly through the PTEN/AKT signaling pathway. PMID: 29901196
- These results suggest that miR214 mediates vascular inflammation and apoptosis via PTEN expression. PMID: 29916551
- A novel information on the susceptibility of PTEN to the inflammatory oxidant HOCl and its effects on the structure and activity of the protein is provided. PMID: 29298524
- The study proposes a new mechanism by which loss of PTEN and consequent activation of the PI3K-AKT-mTORC1-S6K1 signaling pathway impairs DNA repair by downregulation of MRE11. PMID: 28967905
- In prostate tumor tissue microarrays, loss of PTEN phosphohydrolase (PTEN) correlates with increased tyrosine kinase 6 PTK6 tyrosine 342 (PY342) phosphorylation and poor outcome. PMID: 29142193
- In silico analysis revealed PTEN to be the downstream target of miR-21, which was further confirmed by expression analysis. PMID: 29807978
- The decreased PTEN was associated with poorer survival outcomes of patients with kidney cancer, and PTEN acts as a tumor suppressor in tumorigenesis and progression in kidney cancer. PMID: 29408173
- MiR-221, along with proteins MDR1 and ABCG2, was upregulated in cisplatin-resistant A549 lung cancer cells. Anti-miR-221 inhibits proliferation and induces senescence in lung cancer cells. The PTEN/Akt pathway axis was identified as a target of drug resistance induced by miR-221. PMID: 29876362
- These results demonstrate that SPAG6 silencing induces PTEN expression to regulate apoptosis through the PI3K/AKT pathway, indicating that SPAG6 may be a potential therapeutic target for myelodysplastic syndromes. PMID: 29749435
- The inhibition of PTEN also reduced the cancer effects of CD4+ T cells on non-small cell lung cancer (NSCLC) cell lines following miR-142-5p downregulation. Therefore, our study demonstrated that miR-142-5p regulated CD4+ T cells in human NSCLC through PD-L1 expression via the PTEN pathway. PMID: 29767245
- A statistically significant association between PTEN loss and triple negative breast cancers was found in African American women. PMID: 29653745
- miR-130b was upregulated in the lupus nephritis group compared with the control group. PTEN was identified as a virtual target of miR-130b, and there was a negative regulatory association between miR-130b and PTEN. miR-130b and PTEN interfered with the viability and apoptosis of mesangial cells. PMID: 29620214
- The results of the present study indicate that the expression of miRNA23a may regulate acute myocardial infarction (AMI) through targeting PTEN in patients and in vitro, and PTEN/miRNA23a may therefore be potential targets for the clinical treatment of AMI. PMID: 29488607
- TRPC1 regulated HIF1alpha levels in PTEN-deficient MDA-MB-468 and HCC1569 breast cancer cell lines. This regulation arises from effects on the constitutive translation of HIF1alpha under normoxic conditions via an Akt-dependent pathway. PMID: 28559303
- miR367 was revealed to bind directly to phosphatase and tensin homolog (PTEN) mRNA and regulate the expression of the PTEN protein. PMID: 29512776
- The present study confirmed that pAURKA is important in the development of gastric adenocarcinoma and revealed a novel functional link between PTEN, AURKA, and pAURKA activation. PMID: 29512701
- The study found that CKS2 knockdown induced PTEN up-regulation and may associate with P53 pathway activation. PMID: 29487004
- The study showed for the first time that the suppression of rheumatoid arthritis fibroblast-like synoviocyte was mediated by phosphatase and tensin homolog involving survivin silencing. PMID: 28337018
- The overexpression of PTEN concomitant with Livin gene silencing was confirmed as a feasible and effective in vitro and in vivo gene modulation method, which may represent a potential therapeutic strategy for the treatment of gastric cancer. PMID: 29436592
