pdcd10a Antibody

What is PDCD10 and why is it significant in research?

PDCD10, also known as CCM3 (Cerebral Cavernous Malformations 3) and TFAR15, is an evolutionarily conserved protein associated with cell apoptosis. Its significance stems from its role in multiple cellular processes, including regulation of apoptosis, cell proliferation, and vascular development. Research has shown PDCD10 interacts with serine/threonine protein kinase MST4 to modulate the extracellular signal-regulated kinase (ERK) pathway . PDCD10 is of particular interest because mutations in this gene cause cerebral cavernous malformations, which are vascular malformations that can lead to seizures and cerebral hemorrhages . Additionally, PDCD10 has been implicated in cancer development, particularly in osteosarcoma progression .

What are the key characteristics of PDCD10 antibodies?

PDCD10 antibodies are available in both polyclonal and monoclonal formats, with varied host organisms (predominantly rabbit) . These antibodies typically recognize epitopes within the human PDCD10 protein (212 amino acids in length) . The observed molecular weight of PDCD10 is approximately 25-30 kDa . PDCD10 antibodies demonstrate reactivity across multiple species, particularly human, mouse, and rat samples . The antibodies are most commonly used in applications such as Western blotting, immunohistochemistry, immunofluorescence, and immunoprecipitation .

What are the primary research applications for PDCD10 antibodies?

PDCD10 antibodies have been extensively utilized in:

• Cancer research, particularly in osteosarcoma, ovarian, breast, and prostate cancers

• Vascular development studies, especially relating to cerebral cavernous malformations

• Cellular apoptosis pathway investigations

• Protein-protein interaction studies (e.g., with PP2A and ERK pathways)

• Epithelial-mesenchymal transition (EMT) research in tumor progression

How are PDCD10 antibodies validated for research use?

Validation of PDCD10 antibodies typically involves:

• Western blot analysis in multiple cell lines (e.g., MCF-7, PC-3, Raji cells, U2OS, MG63)

• Knockout/knockdown validation through siRNA experiments

• Immunohistochemistry in cancer tissues (e.g., colon cancer)

• Reactivity testing across multiple species samples

• Epitope specificity testing through competitive binding assays

• Verification of nuclear and cytoplasmic localization patterns

What are the optimal conditions for Western blotting with PDCD10 antibodies?

For Western blot applications, PDCD10 antibodies perform optimally under these conditions:

• Recommended dilution range: 1:200-1:2000 for polyclonal antibodies , with 1:500-1:1000 being most commonly used

• Expected band size: ~25-30 kDa

• Positive control samples: MCF-7 cells, PC-3 cells, Raji cells, A-431 cells, K-562 cells, U-251MG cells

• Buffer composition: PBS with 0.02% sodium azide and 50% glycerol at pH 7.3 is recommended for antibody storage

• Loading controls: Standard housekeeping proteins such as actin can be used to normalize PDCD10 expression

What protocols are recommended for immunohistochemistry with PDCD10 antibodies?

For optimal immunohistochemistry results:

• Dilution range: 1:50-1:500

• Antigen retrieval: TE buffer pH 9.0 is suggested; alternatively, citrate buffer pH 6.0 may be used

• Positive tissue controls: Human colon cancer tissue has demonstrated reliable results

• Detection systems: Both chromogenic and fluorescent secondary detection systems have been validated

• Counterstaining: Standard nuclear counterstains (e.g., hematoxylin) are compatible

How can PDCD10 expression be effectively knocked down for functional studies?

Based on published methodologies:

• siRNA approach: PDCD10-specific siRNAs have been successfully used, with non-targeting siRNAs as controls

• Validation of knockdown efficiency: Western blotting at 48 hours post-transfection shows optimal knockdown

• Functional assays: After PDCD10 knockdown, proliferation assays (e.g., CCK-8), migration assays (wound healing), invasion assays (Transwell), and apoptosis assessment (7-AAD labeling) have been successfully employed to determine PDCD10 function

• Expected outcomes: PDCD10 knockdown typically results in inhibited proliferation, migration, and invasion capabilities, along with increased apoptosis in PDCD10-dependent cell lines

How does PDCD10 contribute to tumor progression in osteosarcoma?

Research evidence indicates PDCD10 promotes osteosarcoma progression through multiple mechanisms:

Key pathways affected by PDCD10:

• EMT pathway activation: PDCD10 reduces expression of epithelial markers (e.g., E-cadherin) and transforms cytokeratin into a vimentin-based cytoskeleton

• Apoptosis inhibition: PDCD10 interacts with the Bcl-2 family, caspase family, and potentially p53 to suppress programmed cell death

• Enhanced proliferation: PDCD10 knockdown significantly inhibits osteosarcoma cell proliferation, suggesting its role in promoting cell division

Clinical significance:

• High PDCD10 expression in osteosarcoma patients correlates with worse prognosis

• Positive PDCD10 expression is associated with higher five-year mortality rates

• Bioinformatics analysis of GSE17679 database confirms significant PDCD10 expression differences between normal (n=18) and cancer samples (n=99)

What molecular interactions has PDCD10 been shown to participate in?

PDCD10 engages in several important protein-protein interactions:

These interactions highlight PDCD10's role as a signaling node that integrates multiple cellular pathways involved in survival, proliferation, and vascular development.

How do PDCD10 antibodies contribute to understanding differential PDCD10 expression in various cancer types?

PDCD10 antibodies have revealed variable expression patterns across cancer types:

• Osteosarcoma: 86.84% of patients show positive PDCD10 expression by immunohistochemistry

• T-cell malignancies: Constitutive expression in both non-malignant and malignant T cells, with functional relevance in specific malignant T-cell lines

• Other documented cancer types: Ovarian, breast, prostate, and brain tumors show PDCD10 expression

Research using PDCD10 antibodies has enabled:

• Correlation of expression levels with patient outcomes (e.g., five-year mortality)

• Identification of subcellular localization differences between cancer types

• Evaluation of PDCD10 as a potential therapeutic target, particularly in osteosarcoma

What is the relationship between PDCD10 and the epithelial-mesenchymal transition in cancer progression?

PDCD10 has been identified as a key regulator of EMT:

Mechanistic evidence:

• PDCD10 activates the EMT pathway in osteosarcoma cells

• EMT markers affected by PDCD10 include E-cadherin (decreased) and vimentin-based cytoskeleton development (increased)

• PDCD10 overexpression enhances migration and invasion capabilities of osteosarcoma cells, key functional outcomes of EMT activation

Experimental validation:

• Plate cloning assays, wound healing tests, and Transwell invasion assays all demonstrate enhanced metastatic potential in cells with PDCD10 overexpression

• Animal models confirm increased tumor growth with PDCD10 overexpression

• Western blot analysis confirms altered EMT marker expression profiles dependent on PDCD10 status

What are common challenges when using PDCD10 antibodies and how can they be addressed?

Researchers frequently encounter these challenges when working with PDCD10 antibodies:

How can researchers distinguish between PDCD10 isoforms or post-translationally modified variants?

Recommended approaches:

• Use phospho-specific antibodies when studying PDCD10 phosphorylation by serine/threonine kinase 25

• Employ 2D gel electrophoresis to separate post-translationally modified variants

• Validate observations with mass spectrometry analysis

• Use immunoprecipitation followed by specific post-translational modification antibodies (e.g., phospho, ubiquitin)

• Consider the use of phosphatase treatment to confirm phosphorylation status

How can PDCD10 antibodies be effectively used in combination with other biomarkers?

For comprehensive pathway and functional analyses, PDCD10 antibodies can be combined with:

For apoptosis studies:

• Bcl-2 family proteins (Bcl-2, Bax, etc.)

• Caspase family proteins (Caspase-3, -8, -9)

• p53 for tumor suppressor pathway analysis

For EMT investigation:

• E-cadherin (epithelial marker)

• Vimentin (mesenchymal marker)

• N-cadherin, Snail, and Slug (EMT transcription factors)

For signaling pathway analysis:

• PP2A components

• ERK and phospho-ERK

• MST4 and related kinases

Multiplexed immunofluorescence or sequential immunohistochemistry can be particularly valuable for co-localization studies of these markers.

What emerging applications of PDCD10 antibodies show promise in cancer research?

Several emerging research directions show potential:

• Therapeutic target validation: PDCD10 knockdown inhibits tumor growth, suggesting it could be a viable therapeutic target

• Biomarker development: High PDCD10 expression correlates with poor prognosis in osteosarcoma, indicating potential as a prognostic biomarker

• Combination therapy approaches: Understanding PDCD10's role in apoptosis resistance could inform combination strategies with existing chemotherapeutics

• Metastasis prediction: Given PDCD10's involvement in EMT, antibody-based assays might help predict metastatic potential

How might PDCD10 research intersect with emerging immunotherapy approaches?

While the search results don't directly address PDCD10 and immunotherapy, potential connections exist:

• Recent advancements in PD-1/CTLA-4 bispecific antibodies for cancer therapy suggest potential for exploring PDCD10's role in the tumor immune microenvironment

• PDCD10's apoptosis regulation could affect cancer cell response to immune-mediated cell death

• Investigation of PDCD10 expression in tumor-infiltrating lymphocytes might provide insights into immune evasion mechanisms

• The effect of immune checkpoint inhibitors on PDCD10 expression and function represents an unexplored research area

What additional validation studies would strengthen PDCD10 antibody applications?

To further enhance PDCD10 antibody reliability:

• Expanded knockout/knockdown validation across additional cell types

• Cross-validation using multiple antibody clones targeting different PDCD10 epitopes

• Comprehensive species reactivity profiling beyond human, mouse, and rat

• Spatial proteomics approaches to definitively map PDCD10 subcellular localization

• ChIP-seq applications to identify potential PDCD10 interactions with chromatin