PUB44 Antibody
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Gene References and Functions
- The E3 ligase SAUL1 serves as a positive regulator of PAMP-triggered immunity. Its homeostasis is monitored by the SOC3. PMID: 28691210
- NORE1/SAUL1 is a key factor that integrates signals from temperature-dependent defense programs and leaf senescence in Arabidopsis. PMID: 26910207
- This study demonstrated that Arabidopsis with a mutation of SAUL1 exhibit characteristics of an autoimmune mutant. PMID: 26505534
- The Arabidopsis PUB43 and PUB44 genes are proposed to function during seed germination and early seedling growth. PMID: 21738636
- C-terminal armadillo repeats are essential for SAUL1 localization to the plasma membrane. PMID: 20956359
- SAUL1 represents a regulatory element that prevents senescence from occurring prematurely by targeting AAO3 for degradation. PMID: 19309463
Q&A
Here’s a structured collection of FAQs tailored for academic researchers working with PUB44 antibody, based on experimental methodologies and findings from recent studies:
How does experimental design differ when studying PUB44 in monocots (e.g., rice) vs. dicots (e.g., Arabidopsis)?
Key consideration: Species-specific antibodies must be validated via cross-reactivity tests.
How to resolve contradictions in PUB44-dependent protein degradation kinetics across studies?
Discrepancies arise from:
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Treatment duration: PBI1 degradation in rice peaks at 60–90 min post-chitin , while CPK4 stabilization in Arabidopsis occurs within 24h of NaCl exposure .
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Ligase activity: Recombinant PUB44 shows weak in vitro ubiquitination activity , necessitating in planta assays.
Methodological solution:
What controls are critical for immunoblot quantification of PUB44-mediated degradation?
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Internal standards: Use constitutively expressed proteins (e.g., actin) for normalization.
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Negative controls: Include PUB44-kd cell lines to confirm degradation specificity .
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Proteasome inhibition: Treat with MG132 to validate ubiquitin-proteasome system dependency .
How to address challenges in detecting weak PUB44-substrate interactions?
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Enhanced crosslinkers: Use DSS or EDC/NHS for stabilizing transient interactions.
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Tag optimization: Employ split-luciferase systems (e.g., SmBiT/LgBiT) for sensitive detection in rice protoplasts .
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Phospho-mimetic mutants: Mimic PUB44 phosphorylation (induced by chitin ) to strengthen binding.
What advanced techniques complement PUB44 antibody-based studies?
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Structural analysis: Cryo-EM of PUB44-PBI1 complex to map DUF1110 domain interactions .
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Phosphoproteomics: Identify phosphorylation sites on PUB44 regulating ligase activity.
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Single-cell imaging: Track PUB44-PBI1 dynamics using FRET in live cells.
