unc-51 Antibody
Description
Definition and Biological Context
UNC-51/ULK1 is a serine/threonine kinase that initiates autophagy by phosphorylating downstream substrates like ATG13 and FIP200 . It regulates processes such as bacterial clearance , axonal transport , and mitochondrial homeostasis . Antibodies targeting UNC-51/ULK1 enable researchers to study its expression, post-translational modifications, and interactions in diverse biological systems.
Validation and Performance Data
These antibodies are validated for specificity and sensitivity across species and experimental conditions:
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ULK1 (R600) #4773: Detects endogenous ULK1 in human and monkey cells . Used in studies linking ULK1 to xenophagy and DNA damage-induced autophagy .
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ULK1 (D9D7) #6439: Monoclonal antibody validated for IP and WB, critical for analyzing ULK1 kinase activity in cancer models .
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Abcam ab167139: Recognizes ULK1 in human, mouse, and rat tissues, with applications in neurodevelopmental studies .
Autophagy and Xenophagy
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ULK1 antibodies confirmed its role in bacterial ubiquitylation and p62 recruitment during Listeria monocytogenes clearance .
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Phosphorylation of p62 at S409, mediated by ULK1, enhances ubiquitin affinity and autophagosome formation .
Neuronal Development
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Antibodies revealed ULK1’s interaction with UNC-14 and kinesin motors for axonal transport in Drosophila and C. elegans .
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ULK1 deficiency disrupts synaptic vesicle transport, leading to axonal defects .
Disease Mechanisms
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ULK1 inhibition via RPM-1 ubiquitin ligase activity modulates autophagy in neurodegenerative models .
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Antibodies identified ULK1 as a therapeutic target in cancers, with inhibitors like SBI-0206965 blocking autophagic flux .
Protocol Considerations
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Western Blotting: Use 1:1000 dilution for ULK1 (D9D7) #6439 in RIPA buffer .
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Immunoprecipitation: Optimize lysates with protease/phosphatase inhibitors .
Notable Findings Using UNC-51 Antibodies
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ULK1-p62 Axis: ULK1 phosphorylates p62 at S409 to enhance bacterial targeting .
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Cancer Therapy: ULK1 inhibitors (e.g., MRT68921) synergize with chemotherapy by blocking cytoprotective autophagy .
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Lipotoxicity: ULK1 degrades KEAP1 to activate NFE2L2, mitigating palmitic acid-induced cell death .
Limitations and Challenges
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Cross-Reactivity: Some antibodies may detect ULK2 due to homology .
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Post-Translational Modifications: Phosphorylation or ubiquitination can alter ULK1’s molecular weight, complicating WB interpretation .
Future Directions
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Develop isoform-specific antibodies to distinguish ULK1/ULK2 functions.
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Optimize antibodies for spatial proteomics to map ULK1’s subcellular dynamics.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Mutants with impaired growth cone functions, such as UNC-51, often exhibit abnormal terminations and inappropriate trajectories at the distal ends of the M2 axons. PMID: 12885562
- The autophagy-related kinase UNC-51 regulates the subcellular localization of the Netrin receptor UNC-5 in Caenorhabditis elegans PMID: 16887826
Q&A
Here’s a structured FAQ for researchers working with unc-51 antibody in academic contexts, synthesized from peer-reviewed studies and technical methodologies:
How does UNC-51 kinase activity influence axonal transport vs. autophagy?
Key findings:
Experimental design:
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Use kinase-dead mutants (e.g., unc-51 K38A) to dissect phosphorylation-dependent vs. independent roles .
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Combine autophagy reporters (e.g., LC3-II puncta) with UNC-51 knockdown in neuron-specific models .
How to resolve contradictory data on UNC-51’s role in axon guidance vs. termination?
Case analysis:
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Axon guidance: UNC-51/UNC-14 complexes mediate growth cone dynamics via SynGAP interaction .
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Axon termination: RPM-1 ubiquitinates UNC-51 to inhibit its activity, ensuring proper synapse maturation .
Strategy:
What controls are critical for UNC-51 antibody-based assays in developmental studies?
Best practices:
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Negative controls:
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Positive controls:
How to design a study investigating UNC-51’s dual role in dendrite and axon development?
Integrated approach:
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Model system: Use Drosophila mushroom body neurons, where UNC-51 regulates both dendritic overshooting and axonal Fas II trafficking .
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Tools:
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Readouts:
