PKN3 Antibody, HRP conjugated
Description
Definition and Structure
The PKN3 Antibody, HRP conjugated, is a polyclonal or monoclonal antibody raised against PKN3 and chemically linked to HRP for enhanced detection in assays like Western blot (WB), immunohistochemistry (IHC), and ELISA. Key features include:
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Target: PKN3 (UniProt ID: Q6P5Z2), a 99 kDa protein kinase involved in Rho GTPase signaling and cancer invasiveness .
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Conjugate: HRP enables chromogenic or chemiluminescent signal amplification .
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Host: Typically rabbit-derived, with reactivity across human, mouse, and rat samples .
Applications in Research
PKN3 Antibody, HRP conjugated, is widely used in:
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Western Blot: Detects PKN3 at ~99 kDa in lysates from cancer cell lines (e.g., PC-3, LNCaP) .
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Immunohistochemistry/Immunofluorescence: Localizes PKN3 in tumor tissues, particularly in the nucleus and perinuclear regions .
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Flow Cytometry: Identifies PKN3 expression in live or fixed cells (e.g., prostate carcinoma cells) .
Role in Cancer Signaling
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PKN3-p130Cas Interaction: PKN3 binds and phosphorylates p130Cas, an adaptor protein critical for tumor invasiveness. This interaction occurs via the SH3 domain of p130Cas and a polyproline motif in PKN3 .
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Phosphorylation Sites: PKN3 phosphorylates p130Cas at Ser428/432, correlating with enhanced cell migration and tumor growth in breast and prostate cancers .
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Angiogenesis and Metastasis: PKN3 knockout mice show impaired angiogenesis and reduced lung metastasis in melanoma models, linked to defective glycosylation of cell-surface proteins (e.g., integrins) .
Table 2: Key Phosphorylation Sites in PKN3 (Q6P5Z2)7
Technical Considerations
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Buffer Compatibility: HRP conjugation efficiency depends on antibody buffer composition; additives like sodium azide or BSA may interfere .
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Validation: Knockout-validated antibodies (e.g., Novus Biologicals NBP130102H) ensure specificity .
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Cross-Reactivity: Predictions suggest reactivity with pig, bovine, and dog homologs due to sequence homology .
Clinical and Preclinical Relevance
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Steady-state kinetic analysis has shown that PKN1-3 follows a sequential ordered Bi-Bi kinetic mechanism, where peptide substrate binding occurs before ATP binding. This kinetic mechanism has been further confirmed by additional kinetic studies focusing on product inhibition and affinity of small molecule inhibitors. PMID: 27919031
- PKN3 plays a major role in regulating angiogenesis in humans and mice, and tumor metastasis in mice. PMID: 26742562
- PKN3 has been identified as a novel protein involved in remodeling the actin-adherens junction, potentially by linking ICAM-1-signaling with actin/AJ dynamics. PMID: 22609186
- PKN isoforms are not merely redundant in supporting migration but appear to be interconnected through isoform-specific regulatory domain properties, linked to selective upstream signals. PMID: 21754995
- PKN3 may represent a promising target for therapeutic intervention in cancers lacking tumor suppressor PTEN function or those dependent on chronic activation of phosphoinositide 3-kinase. PMID: 15282551
