par-6 Antibody
Description
Target Protein Profile
Par6 exists as three isoforms (α, β, γ) encoded by PARD6A, PARD6B, and PARD6G genes respectively. Key features include:
Cancer Mechanism Studies
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Breast Cancer: Par6α antibodies detect overexpression in 68% of ductal carcinoma in situ lesions and 82% of invasive carcinomas
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Ovarian Cancer: IHC with Par6 antibodies revealed 3.2x higher expression in metastatic vs primary tumors (p<0.01)
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EMT Regulation: Co-staining with E-cadherin/Vimentin shows Par6's role in TGFβ-induced epithelial-mesenchymal transition
Subcellular Localization Analysis
Nuclear fractionation studies using Par6 antibodies demonstrated:
| Cell Line | Cytoplasmic (%) | Nuclear (%) | Detection Method |
|---|---|---|---|
| MDCK | 92 | 8 | Subcellular fractionation |
| HeLa | 60 | 40 | Immunofluorescence |
Antibody: Affinity-purified rabbit polyclonal (GST-Par6 C-terminus)
Technical Validation Data
Critical performance parameters from recent studies:
Clinical Correlation Findings
Par6 antibody-based studies have revealed significant disease associations:
Functional Studies
Par6 antibodies enabled critical mechanistic insights through:
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Domain Mapping: Truncation mutants identified Cdc42-binding domain (ΔPro136) as essential for proliferation induction
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Pathway Analysis: Co-IP experiments revealed Par6-aPKC-Cdc42 complex activates MEK/Erk signaling (1.8x phospho-Erk increase)
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Metastasis Models: shRNA knockdown reduced lung metastases by 67% in xenografts (p<0.01)
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research has shown that vab-1, along with CDC-42, is essential for polarizing CDC-42 activity towards the medial tip during epithelial intercalation. This discovery reveals a previously unidentified role for polarized CDC-42, working in conjunction with PAR-6, PAR-3, and an Eph receptor (vab-1), during epithelial intercalation. PMID: 27861585
- The arcade cell epithelium exhibits polarization through a PAR-6-mediated pathway that operates independently of E-cadherin, beta-integrin, and beta-laminin. PMID: 25773364
- PAR-3 and PAR-6 demonstrate a degree of redundancy in their functions during cytokinesis, contributing to the maintenance of DYN-1 within the cleavage furrow. PMID: 22887994
- Evidence suggests that PAR polarity arises from the coupling of advective transport driven by flowing cell cortex with a multistable PAR reaction-diffusion system. Advection within an active, flowing medium provides a mechanism for mechanically templated pattern formation during development. PMID: 22021673
- Centrosomes achieve apical localization by first moving towards lateral foci of PAR-3 and PAR-6, conserved polarity proteins. Subsequently, they move together with these foci towards the future apical surface. Notably, embryos lacking PAR-3 fail to localize their centrosomes apically. PMID: 22425160
- PAR-2 and PAR-6, residing in opposing PAR domains, undergo exchange between well-mixed cytoplasmic populations and laterally diffusing membrane-associated states. PMID: 21518794
- Binding to PKC-3, but not to PAR-3 or a conventional PDZ domain ligand, is crucial for PAR-6 function in C. elegans. PMID: 20122916
- PAR proteins play roles in both apicobasal and anterior-posterior asymmetry during the first few cell cycles of embryogenesis. PMID: 13129846
- CDC-37-mediated inhibition of the CDC-42-dependent binding site and PAR-3-mediated release of this inhibition constitute a key mechanism for the anterior accumulation of PAR-6. PMID: 16943281
- CDC-42 interaction with PAR-6 is not essential for the initial establishment of asymmetry, but it is required for maximal cortical accumulation of PAR-6 and for maintaining its asymmetry. PMID: 16996049
- PAR-6 is indispensable for junction formation in C. elegans embryonic epithelial cells, but not for apicobasal polarization. PMID: 17314130
- Loss of function mutations in nos-3 suppress the lethality associated with par-2 mutations by regulating PAR-6 protein levels. PMID: 18502413
- Research has identified a RhoGAP, PAC-1, which mediates embryo radial polarity and gastrulation by excluding PAR-6 from contacted cell surfaces. PAC-1 serves as a dynamic molecular link between cell contacts and PAR proteins, contributing to the radial polarization of embryos. PMID: 18583611
Q&A
Here’s a structured FAQ collection tailored to academic research on Par-6 antibodies, derived from peer-reviewed studies and technical documentation:
How to validate Par-6 antibody specificity in immunofluorescence?
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Methodology:
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Preabsorption controls: Incubate the antibody with its immunogen (e.g., GST-Par-6 C-terminal fusion protein) and compare staining patterns in cell lines like MDCK or HeLa. Loss of signal confirms specificity .
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Knockdown/knockout validation: Use siRNA or CRISPR to reduce Par-6 expression and verify diminished antibody signal .
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Epitope mapping: Compare antibodies targeting distinct regions (e.g., N-terminal vs. internal regions) to confirm consistent localization .
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What experimental designs optimize Par-6 detection in membrane vs. nuclear compartments?
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Approach:
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Fractionation: Separate cytoplasmic, membrane, and nuclear fractions via differential centrifugation, followed by Western blot (WB) .
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Antibody selection: Use antibodies against membrane-associated domains (e.g., PDZ domain) for junctional staining or nuclear-localizing epitopes .
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Fixation: For nuclear Par-6, use methanol fixation over paraformaldehyde to preserve epitopes .
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Which applications are Par-6 antibodies most suitable for?
How to resolve contradictory Par-6 localization data across studies?
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Troubleshooting:
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Antibody cross-reactivity: Test across Par-6 isoforms (α/β/γ) using isoform-specific knockdowns .
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Epitope accessibility: Compare staining in permeabilized vs. non-permeabilized cells to assess membrane-bound vs. cytoplasmic pools .
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Context-dependent localization: Assess cell confluency (e.g., polarized MDCK monolayers vs. subconfluent HeLa cells) .
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What functional assays confirm Par-6 antibody efficacy in blocking protein activity?
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Functional validation:
How to address cross-species reactivity limitations in non-model organisms?
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Strategies:
How do Par-6 antibody performance metrics vary by target epitope?
How to design multiplex experiments with Par-6 antibodies?
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Multiplexing guidelines:
