plk-3 Antibody
Description
Definition and Biological Significance
PLK3, also known as CNK or PRK, belongs to the Polo-like kinase family and regulates critical processes such as:
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Cell cycle progression: Required for G1/S phase transition and cyclin E regulation .
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Stress responses: Activated under hypoxia, oxidative stress, or DNA damage to modulate HIF-1α stability and apoptosis .
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Tumor suppression: Low PLK3 expression correlates with malignancies in lung, colon, and pancreatic cancers .
PLK3 antibodies target specific epitopes of the 72 kDa protein, facilitating its detection in human, mouse, and rat samples .
Key Applications of PLK3 Antibodies
Research Findings Using PLK3 Antibodies
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Hypoxia and Tumor Angiogenesis:
PLK3 destabilizes HIF-1α under hypoxic conditions, reducing VEGF-A expression and tumor vascularization . PLK3-null mice exhibit increased tumor blood vessel density . -
Cell Cycle Regulation:
PLK3 depletion arrests cells in G1 phase by suppressing cyclin E and Cdc25A, critical for S-phase entry . -
Pancreatic Cancer:
PLK3 loss in PDAC (pancreatic ductal adenocarcinoma) promotes metastasis by inhibiting anoikis, a caspase-dependent apoptosis mechanism . -
Oxidative Stress:
PLK3 knockdown enhances antioxidant capacity (e.g., SOD, GSH-Px) in Leydig cells, mitigating H2O2-induced senescence .
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
How do researchers validate PLK3 antibody specificity for immunoprecipitation (IP) and Western blot (WB) applications?
Basic Research Focus
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Knockout validation: Use PLK3−/− cell lines or siRNA-mediated knockdown controls to confirm signal absence in WB/IP experiments .
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Epitope mapping: Compare antibody performance against truncated PLK3 domains (e.g., NT1, CT1) to identify recognized regions (Figure S2E in ).
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Cross-reactivity testing: Validate against other PLK family members (e.g., PLK1/PLK2) using lysates from cells overexpressing these kinases .
What experimental strategies resolve contradictory subcellular localization data for PLK3 isoforms?
Advanced Research Focus
PLK3 exhibits isoform-specific localization influenced by antibody epitopes:
| Antibody Clone | Epitope Region | Localization Observed | Key Reference |
|---|---|---|---|
| Abcam (Polyclonal) | N-terminal (NT1/NT2) | Membrane/nuclear-cytoskeletal | |
| BD Biosciences (Monoclonal) | Kinase-PBD linker | Cytoplasmic |
Methodological Solutions:
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Perform subcellular fractionation (cytoplasmic/membrane/nuclear) with separation validated by marker proteins (e.g., Lamin B1 for nuclear fractions) .
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Combine PLA (Proximity Ligation Assay) with isoform-specific antibodies to map spatial interactions (e.g., PLA confirmed CD95-Plk3 proximity in HeLa membranes) .
How can phosphorylation-dependent PLK3 activation be distinguished from isoform migration artifacts?
Advanced Research Focus
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Lambda phosphatase treatment: Incubate lysates with phosphatase to eliminate phosphorylation-induced mobility shifts (Figure S2C in ).
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SILAC mass spectrometry: Use heavy/light isotope labeling during IP-MS to differentiate phosphorylation states (Table S1 in ).
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Kinase-dead mutants: Compare migration patterns of wild-type vs. K52R (kinase-dead) PLK3 in overexpression systems .
What controls are essential for PLK3 immunohistochemistry (IHC) in cancer tissue studies?
Basic Research Focus
Validation: Correlate IHC results with WB using microdissected tumor regions .
How do researchers analyze PLK3’s role in apoptosis signaling pathways?
Advanced Research Focus
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Kinase activity assays: Measure PLK3-mediated phosphorylation of caspase-8 (T273) via in vitro kinase assays with recombinant proteins .
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Time-course stimulation: Treat cells with CD95L and track PLK3-caspase-8 interaction decay via co-IP (Figure 2F in ).
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Pathway inhibition: Use PLK3 inhibitors (e.g., GW843682X) or dominant-negative mutants to block apoptosis in CD95-stimulated cells .
What in vivo models are suitable for studying PLK3’s tumor-suppressive functions?
Advanced Research Focus
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PLK3 knockout mice: Monitor spontaneous tumor development (e.g., increased lung adenocarcinoma incidence in Plk3−/− mice) .
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Xenograft models: Compare tumor growth in WT vs. PLK3-silenced cell lines under hypoxic conditions .
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Angiogenesis assays: Quantify blood vessel density (via vWF staining) in Plk3−/− tumors (Figure 5 in ).
How are PLK3 antibody performance differences addressed in stress-response studies?
Methodological Recommendations:
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Antibody cross-validation: Compare multiple clones (e.g., Proteintech 10977-1-AP vs. R&D Systems AF4197) in parallel WB/IP experiments .
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Stress condition optimization: Adjust lysis buffers to preserve post-translational modifications (e.g., 1% NP-40 + phosphatase inhibitors for DNA damage studies) .
