PTPN6 Antibody
Product Specs
Target Background
- This study provides compelling in vivo and cellular evidence that hepatocyte SHP-1 plays a pivotal role in the production of inflammatory mediators that contribute to endotoxemia. PMID: 28533521
- Actomyosin retrograde flow regulates the immune response of primary human Natural Killer cells through a novel interaction between beta-actin and SHP-1, altering its conformational state and thereby regulating Natural Killer cell cytotoxicity. PMID: 29449322
- Biophysical assays of tethered signaling reactions demonstrate that tether-controlled activity is a key aspect of the phosphatase SHP-1. PMID: 28378014
- This research demonstrates that VB inhibits glioblastoma cell proliferation, migration, and invasion while promoting apoptosis through SHP-1 activation and inhibition of STAT3 phosphorylation. PMID: 29961065
- Mycobacterium tuberculosis-initiated human mannose receptor signaling regulates macrophage recognition and vesicle trafficking through gamma Fc receptors, Grb2, and SHP-1. PMID: 28978467
- Data suggests that the SHP-1/p-STAT3/VEGF-A axis is a potential therapeutic target for metastatic triple-negative breast cancer (TNBC). PMID: 27364975
- These findings highlight a signaling pathway in which SHP-1 acts through CrkII to reshape the pattern of Rap1 activation in the immunological synapse. PMID: 28790195
- Observations suggest that Chikungunya virus (CHIKV) has the ability to induce host PTPN6 expression, and induction of PTPN6 may favor the attenuation of the pro-inflammatory immune response of the host, which is otherwise detrimental for the survival of CHIKV and the establishment of an infection. PMID: 29058147
- The results reveal that SHP1 is the long-sought phosphatase that can antagonize Helicobacter pylori CagA. Increased Helicobacter pylori CagA activity, via SHP1 inhibition, might also contribute to the development of Epstein-Barr virus-positive gastric cancer. PMID: 27572445
- Analysis of gene expression profiles of monocytes from symptomatic congestive heart failure patients shows a down-regulation of the phosphatase SHP-1, which induces a significant activation of TAK-1/IKK/NF-kB signaling. PMID: 27814644
- Crocin induced the expression of SHP-1, a tyrosine protein phosphatase, and pervanadate treatment reversed the crocin-induced downregulation of STAT3, suggesting the involvement of a protein tyrosine phosphatase. PMID: 28295507
- This review focuses on the implication of SHP-1 in the pathogenesis of autoimmune disorders and addresses developing therapeutic strategies targeting SHP-1. PMID: 27216862
- We demonstrated that SHP-1 dephosphorylates PKM2Y105 to inhibit the Warburg effect and nucleus-dependent cell proliferation. The dephosphorylation of PKM2Y105 by SHP-1 determines the efficacy of targeted drugs for hepatocellular carcinoma treatment. PMID: 26959741
- These findings provide initial evidence that PDZK1 expression is negatively correlated with SHP-1 activation and poor clinical outcomes in clear cell renal cell carcinoma (ccRCC). PDZK1 was identified as a novel tumor suppressor in ccRCC by negating SHP-1 activity. PMID: 28692056
- Luteolin inhibited STAT3 activation by disrupting the binding of HSP-90 to STAT3, which promoted its interaction with SHP-1. PMID: 28182003
- These findings show a novel role for Shp-1 in the regulation of IEC growth and secretory lineage allocation, possibly through modulation of PI3K/Akt-dependent signaling pathways. PMID: 28465325
- The role of Shp1 in myeloid cells and how its dysregulation affects immune function, which can impact human disease, is reviewed. PMID: 28606940
- PTPN6 is associated with the progression of chronic myeloid leukemia. Low expression levels of PTPN6 were associated with DNA methylation and regulated by histone acetylation. PMID: 28480959
- Shp1 functions as a positive regulator and acts through a novel mechanism by promoting EGFR protein expression in human epithelial cells. PMID: 28416389
- SHP1 DNA methylation is investigated in patients with B cell non-Hodgkin lymphoma. PMID: 28210822
- These results indicate that DNMT1 mediates aberrant methylation and silencing of the SHP-1 gene in chronic myelogenous leukemia cells. PMID: 28376405
- Results provide evidence that repression of SHP-1, SHP-2 and SOCS-1 gene expression in patient plasma cells supports the constitutive activation of the JAK/STAT3 pathway and likely leads to higher degrees of bone marrow invasion. PMID: 28369102
- We found that THEMIS directly regulates the catalytic activity of the tyrosine phosphatase SHP-1. PMID: 28250424
- Hyperglycemia induces SHP-1 promoter epigenetic modifications, causing its persistent expression and activity, leading to insulin resistance, podocyte dysfunction, and diabetic nephropathy (DN). PMID: 27585521
- Low SHP1 expression is associated with primary central nervous system lymphoma. PMID: 27959415
- This study evaluated SHP1-P2 methylation levels in the lymph nodes of colorectal cancer (CRC) patients. It was hypothesized that SHP1-P2 methylation levels would be higher in metastatic lymph nodes. PMID: 27644671
- Overexpression of SHP1 downregulates the JAK2/STAT3 pathway to modulate various target genes and inhibit cell proliferation, migration, and invasion in gastric cancer cells. PMID: 26508024
- This study also found no correlation of SHP-1 expression at diagnosis with response to treatment, although a trend for lower SHP-1 expression was noted in the very small non-responders' group of the 3-month therapeutic milestone. PMID: 26373709
- SYK, LYN and PTPN6 were markedly elevated in atherosclerotic plaques of carotid atherosclerosis patients. PMID: 26742467
- cAMP signaling of Bordetella adenylate cyclase toxin through the SHP-1 phosphatase activates the BimEL-Bax pro-apoptotic cascade in phagocytes. PMID: 26334669
- Quinalizarin enhances radiosensitivity of nasopharyngeal carcinoma cells partially by suppressing SHP-1 expression. PMID: 26781335
- N225K and A550V PTPN6 mutations cause loss-of-function leading to JAK3 mediated deregulation of the STAT3 pathway and uncover a mechanism that tumor cells can use to control PTPN6 substrate specificity. PMID: 26565811
- In addition to their role in NK cell activation by hematopoietic cells, the SLAM-SAP-SHP1 pathways influence responsiveness toward nonhematopoietic targets by a process akin to NK cell 'education'. PMID: 26878112
- A combination of sorafenib and SC-43 is a synergistic SHP-1 agonist duo that reduces tumor size and prolongs survival time. PMID: 26679051
- SHP-1 has a critical role in radioresistance, cell cycle progression, and senescence of nasopharyngeal carcinoma cells. PMID: 26215037
- Data demonstrate for the first time that SHP1 methylation has high specificity for the diagnosis of endometrial carcinoma, while CDH13 promoter methylation plays a role in the earlier stage. PMID: 26597461
- MiR-378g enhanced radiosensitivity partially by targeting SHP-1 in NPC cells. PMID: 26473472
- Soluble egg antigens glycans are essential for the induction of enhanced SOCS1 and SHP1 levels in dendritic cells via the mannose receptor. PMID: 25897665
- miR-4649-3p is downregulated in nasopharyngeal carcinoma cell lines accompanied by SHP-1 upregulation. PMID: 26081980
- Results show that dissociation of SHP-1 from spinophilin is followed by an increase in the binding of spinophilin to PP1. PMID: 25785436
- Data show that SHP-1 promotes HIF-1alpha degradation under hypoxic conditions, leading to a reduction in VEGF synthesis and secretion and impairing epithelial cell proliferation. PMID: 25799543
- Phosphorylation of ATR and CHK1 did not show significant differences in the three cell groups. Overexpression of SHP-1 in the CNE-2 cells led to radioresistance, and the radioresistance was related to enhanced DNA DSB repair. PMID: 25962492
- SHP-1 is a potent suppressor of epithelial-mesenchymal transition and metastasis in hepatocellular carcinoma. PMID: 25619838
- Inactivation of SHP1 is associated with myeloproliferative neoplasm. PMID: 25824741
- Data suggest that protein tyrosine phosphatase non-receptor type 6 (SHP-1) may interact with EGF receptor (EGFR) to inhibit proliferation. PMID: 22797910
- SHP-1 expression was lower in PBMCs from unmedicated schizophrenics. The promoter region was hypermethylated. Silencing SHP-1 induced IKK/NF-kB and pro-inflammatory cytokines. SHP-1 expression may explain 30% of the clinical negative symptom variance. PMID: 24793756
- SHP-1(I) presented concordance between unmethylated promoter region and tumor for breast or prostate. PMID: 25635370
- A high level of SHP1P2 methylation of hilar lymph nodes from stage I NSCLC patients is associated with early relapse of disease. PMID: 23824557
- Transcription factor RFX-1 regulates SC-2001-mediated SHP-1 Phosphatase transcription in hepatocellular carcinoma cells. PMID: 24952874
- Decreased expression levels of SHP-1 caused by aberrant promoter hypermethylation may play a key role in the progression of CML by dysregulating BCR-ABL1, AKT, MAPK, MYC, and JAK2/STAT5 signaling. PMID: 24647617
Q&A
What is PTPN6 and what is its significance in research?
PTPN6, also designated as SHP-1, is a non-receptor protein tyrosine phosphatase primarily expressed in hematopoietic cells. It is characterized by two SH2 domains located N-terminal to the PTP domain and maps to chromosome 12p13, a region commonly involved in leukemia-associated chromosomal abnormalities. The protein plays a crucial role in modulating intracellular signaling for various molecules and has been implicated in the control of tyrosine kinase signaling pathways in cellular proliferation, with potential roles in cancer development .
In various malignancies including T-cell lymphomas, SHP-1 phosphatase expression is often lost through mechanisms that remain to be fully elucidated. This loss of expression may contribute to dysregulated signaling pathways that promote cancer progression, making PTPN6 an important target for cancer research .
What are the molecular characteristics of PTPN6 protein?
PTPN6 consists of 597 amino acids with a calculated molecular weight of 68 kDa, though it is typically observed at approximately 63 kDa in gel electrophoresis . This discrepancy between calculated and observed molecular weights may be attributed to post-translational modifications or protein folding effects on gel migration.
The protein is a member of the non-receptor protein tyrosine phosphatase subfamily, characterized by a catalytic phosphatase domain. SHP-1 is cytosolic and has high substrate specificity for phosphotyrosyl proteins, structurally distinct from protein serine phosphatases, protein threonine phosphatases, or acid and alkaline phosphatases .
What types of PTPN6 antibodies are currently available for research?
Researchers have access to several types of PTPN6 antibodies:
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Monoclonal antibodies: Mouse-derived monoclonal antibodies such as CPTC-PTPN6-2 are available for highly specific detection .
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Polyclonal antibodies: Rabbit polyclonal antibodies like AP02795PU-N recognize epitopes around specific phosphorylation sites, such as tyrosine 536 (S-E-Y-G-N) in human SHP-1 .
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Conjugated antibodies: Primary antibodies conjugated with fluorescent dyes including CF®405S, CF®488A, CF®568, CF®594, and CF®640R for various detection methods .
Each antibody type offers distinct advantages depending on the specific research application, with monoclonals providing high specificity and polyclonals offering broader epitope recognition.
How should researchers select the appropriate fluorescent conjugate for PTPN6 detection?
Selection of fluorescent conjugates should be based on the detection platform, target abundance, and potential for background interference. The following table summarizes available conjugates and their applications:
| Antibody Prefix | Conjugation | Excitation/Emission (nm) | Laser Line | Detection Channel | Applications |
|---|---|---|---|---|---|
| BNC04 | CF®405S | 404/431 | 405 | DAPI, AF405 | Microscopy, low background needed |
| BNC88 | CF®488A | 490/515 | 488 | GFP, FITC | Standard fluorescence, good signal |
| BNC68 | CF®568 | 562/583 | 532, 561 | RFP, TRITC | Reduced autofluorescence |
| BNC94 | CF®594 | 593/614 | 561 | Texas Red® | High sensitivity detection |
| BNC40 | CF®640R | 642/662 | 633, 635 | Cy5 | Far-red detection, minimal overlap |
Important consideration: Blue fluorescent dyes like CF®405S are not recommended for detecting low abundance targets due to their lower fluorescence intensity and potentially higher non-specific background compared to other dye colors .
What are the recommended dilutions and applications for PTPN6 antibodies?
Based on validated protocols, PTPN6 antibodies can be used in multiple applications with the following recommended dilutions:
| Application | Recommended Dilution | Notes |
|---|---|---|
| Western Blot (WB) | 1:5000-1:50000 | High sensitivity detection |
| Immunoprecipitation (IP) | 0.5-4.0 μg for 1.0-3.0 mg lysate | Optimal for protein interaction studies |
| Immunohistochemistry (IHC) | 1:50-1:500 | Tissue-dependent optimization required |
| Immunofluorescence (IF/ICC) | 1:50-1:500 | Cell type may influence optimal dilution |
| Flow Cytometry (Intracellular) | 0.20 μg per 10^6 cells in 100 μl | For quantitative expression analysis |
It is strongly recommended to titrate antibodies in each specific experimental system to obtain optimal results, as performance can be sample-dependent .
What cell lines and tissues have been validated for PTPN6 antibody applications?
PTPN6 antibodies have been validated in several experimental systems:
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Western blot: Positive detection in Raji cells, Jurkat cells, and HuH-7 cells
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Immunoprecipitation: Successfully used in Jurkat cells
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Immunohistochemistry: Validated in human colon cancer tissue
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Immunofluorescence/ICC: Positive detection in MCF-7 cells
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Flow Cytometry: Successfully used for intracellular staining in MCF-7 cells
Additionally, PTPN6 expression has been studied in bladder cancer tissues and various colon cancer cell lines including SW480 and LOVO .
What are the optimal buffer conditions for PTPN6 antibody use in IHC?
For immunohistochemical applications, antigen retrieval conditions significantly impact staining quality. The recommended buffer conditions for PTPN6 antibody in IHC applications are:
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Primary recommendation: TE buffer pH 9.0
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Alternative method: Citrate buffer pH 6.0
The choice between these buffer conditions may depend on tissue type, fixation method, and specific epitope accessibility. For human colon cancer tissue in particular, TE buffer at pH 9.0 has been validated to produce optimal results .
How is PTPN6 expression altered in different cancer types?
Research has revealed distinct PTPN6 expression patterns across cancer types:
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Bladder Cancer: PTPN6 is overexpressed in bladder cancer tissues compared to normal bladder tissues, with significant correlation to tumor grade, stage, T classification, and N classification .
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Colon Cancer: PTPN6 expression has been detected in various colon cancer cell lines. Functional studies demonstrate that PTPN6 knockdown significantly inhibits proliferation, migration, and invasion capabilities of colon cancer cells (SW480 and LOVO), suggesting PTPN6 may promote colon cancer progression .
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Lymphomas: Preliminary studies indicate PTPN6/SHP-1 may help differentiate mantle/marginal zone lymphomas (SHP-1 positive) from follicular lymphomas (SHP-1 negative), suggesting potential diagnostic utility .
These varied expression patterns highlight the context-dependent role of PTPN6 in different cancer types.
How does PTPN6 relate to epithelial-mesenchymal transition in cancer?
Research on colon cancer cells demonstrates that PTPN6 may regulate epithelial-mesenchymal transition (EMT), a critical process in cancer metastasis:
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EMT marker regulation: PTPN6 knockdown promotes expression of epithelial marker E-cadherin while inhibiting mesenchymal markers Vimentin and N-cadherin
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Transcription factor modulation: PTPN6 silencing suppresses expression of EMT-associated transcription factors Twist1, TWIST2, ZEB1, and ZEB2
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Matrix metalloproteinase inhibition: Expression levels of MMP2 and MMP9, which facilitate invasion, decrease after PTPN6 silencing in LOVO and SW480 cells
These findings suggest PTPN6 may facilitate cancer progression by promoting EMT and associated invasive properties.
What is the relationship between PTPN6 and immune cell infiltration in tumors?
Analysis of PTPN6 expression in bladder cancer revealed significant associations with immune cell infiltration in the tumor microenvironment:
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Positive correlations were observed with:
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B cells (P = 5.078e-39)
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CD4+ T cells (P = 8.716e-14)
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Dendritic cells (P = 0.003)
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Neutrophils (P = 0.004)
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Negative correlations were found with:
These findings suggest PTPN6 may play a regulatory role in the tumor immune microenvironment, potentially affecting immunosurveillance and response to immunotherapies.
How can phospho-specific PTPN6 antibodies be used to investigate signaling mechanisms?
Phospho-specific antibodies targeting sites like tyrosine 536 in PTPN6 enable detailed investigation of phosphorylation-dependent regulation mechanisms:
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Phosphorylation status monitoring: Antibodies recognizing the phosphorylated form (e.g., AP02795PU-N targeting phospho-Y536) allow researchers to track activation status under different stimuli or in various disease states .
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Signaling pathway analysis: By examining correlations between PTPN6 phosphorylation and downstream effector activation, researchers can map signaling cascades involving this phosphatase.
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Structure-function studies: Comparing total PTPN6 levels versus phosphorylated forms helps elucidate how posttranslational modifications affect enzymatic activity and protein interactions.
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Therapeutic response monitoring: Changes in PTPN6 phosphorylation patterns following drug treatment may serve as pharmacodynamic biomarkers.
What experimental design considerations are important when conducting PTPN6 knockdown studies?
Based on published methodologies, successful PTPN6 knockdown experiments should incorporate:
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Appropriate controls: Include non-targeting siRNA or empty vector controls alongside PTPN6-targeting constructs in all experiments.
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Knockdown verification: Confirm reduced PTPN6 expression at both mRNA level (via qPCR) and protein level (via Western blot) before interpreting phenotypic outcomes.
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Multiple cell lines: Test effects in multiple relevant cell lines (e.g., both SW480 and LOVO for colon cancer studies) to ensure robustness of findings.
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Functional assays: Employ multiple complementary assays (proliferation, migration, invasion) to comprehensively characterize phenotypic consequences.
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Molecular mechanism investigation: Assess changes in relevant downstream pathways (e.g., EMT markers, transcription factors) to elucidate mechanistic insights .
How can co-immunoprecipitation with PTPN6 antibodies reveal novel protein interactions?
Co-immunoprecipitation (Co-IP) using PTPN6 antibodies allows researchers to identify and characterize protein interaction networks:
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Binding partner identification: Using PTPN6 antibodies to pull down protein complexes followed by mass spectrometry can reveal novel interaction partners.
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Interaction domain mapping: By comparing Co-IP results using antibodies targeting different PTPN6 domains, researchers can determine which regions mediate specific protein interactions.
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Stimulus-dependent interactions: Performing Co-IP under various cellular conditions can reveal dynamic, context-dependent protein associations.
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Phosphorylation-dependent interactions: Comparing results using phospho-specific versus pan-PTPN6 antibodies can identify interactions regulated by phosphorylation status.
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Therapeutic target validation: Co-IP studies can validate whether candidate drugs effectively disrupt pathological protein interactions involving PTPN6 .
What are the optimal storage conditions for PTPN6 antibodies?
To maintain antibody integrity and performance, follow these storage guidelines:
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Temperature: Store PTPN6 antibodies at -20°C for long-term stability.
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Aliquoting: Divide antibody solutions into small aliquots to avoid repeated freeze-thaw cycles, which can degrade antibody quality.
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Buffer composition: Most PTPN6 antibodies are supplied in PBS with 0.02% sodium azide and 50% glycerol at pH 7.3, which helps maintain stability during freezing.
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Shipping conditions: When receiving new antibodies, verify they were shipped on blue ice and promptly store at recommended temperatures.
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Shelf life: PTPN6 antibodies typically remain stable for one year after shipment when stored properly .
