SLAC1 Antibody
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Our research, examining the locus of the SLAC1 gene, suggests that this gene, closely associated with stomatal closure, is likely responsible for the observed differences in drought tolerance. PMID: 25695335
- SLAC1 is indispensable for rapid stomatal closure in response to ozone exposure, reduced air humidity, and darkness. PMID: 23703845
- The predictive power of a systems dynamic model in Arabidopsis is demonstrated by its ability to explain the paradoxical suppression of channels facilitating K+ uptake, slowing stomatal opening, due to mutations in the SLAC1 anion channel. PMID: 23090586
- Guard cells with mutations in SLAC1 exhibit significantly reduced inwardly-rectifying K+ channel currents. Decreasing the cytosolic Ca2+ concentration rapidly restores K+ channel currents in SLAC1-mutant guard cells. PMID: 23126621
- Evidence suggests that SLAC1 may serve as a pathway for providing CO2 for photosynthesis in guard cells. PMID: 21931667
- The guard cell anion channel SLAC1 is regulated by CDPK protein kinases with distinct Ca2+ affinities. PMID: 20385816
- SLAC1 represents the slow, deactivating, weak voltage-dependent anion channel of guard cells, controlled by phosphorylation/dephosphorylation mechanisms. PMID: 19955405
- Our research findings indicate that OZS1 contributes to stomatal closure, but it is not involved in responses to light, abscisic acid, high concentrations of carbon dioxide, and ozone. PMID: 18084014
- The loss-of-function mutation in SLAC1 resulted in an over-accumulation of osmoregulatory anions in guard cell protoplasts. PMID: 18305482
- The plasma membrane protein SLAC1 is essential for stomatal closure in response to CO2, abscisic acid, ozone, light/dark transitions, humidity change, calcium ions, hydrogen peroxide, and nitric oxide. PMID: 18305484
Q&A
Here’s a structured collection of FAQs tailored for academic researchers working with SLAC1 antibodies, organized by complexity and methodological focus:
Advanced Research Questions
How can researchers resolve contradictory data on SLAC1 activation mechanisms in calcium-dependent vs. calcium-independent pathways?
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Experimental design:
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Data interpretation: Use a signaling pathway table to contextualize results:
| Pathway | Key Regulators | SLAC1 Current Amplitude (nA) | Citation |
|---|---|---|---|
| Ca²⁺-independent | OST1, ABI1 | 450 ± 60 | |
| Ca²⁺-dependent | CPK21, CPK23 | 320 ± 45 | |
| Phosphomimetic | S59D/S107D/S152D | 620 ± 80 |
What structural insights from cryo-EM studies inform SLAC1 antibody epitope selection?
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Target non-conserved extracellular loops (e.g., residues 105–112 in the latch domain) to avoid cross-reactivity with homologs like SLAH3 .
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Avoid epitopes near phosphorylation sites (e.g., S59, S107, S152) to prevent antibody interference with kinase assays .
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Use surface plasmon resonance (SPR) to quantify antibody binding affinity to SLAC1 extracellular domains (KD < 1 nM recommended) .
How to troubleshoot nonspecific banding in SLAC1 western blots during ABA signaling experiments?
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Step 1: Optimize membrane solubilization with 2% (w/v) digitonin to preserve SLAC1-kinase complexes .
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Step 2: Employ 2D gel electrophoresis to separate SLAC1 isoforms by charge (phosphorylation alters pI by ~0.5 units) .
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Step 3: Validate with kinase inhibitor assays (e.g., 10 μM staurosporine) to confirm phosphorylation-dependent mobility shifts .
Data Contradiction Analysis
Conflicting reports on SLAC1 inhibition by ABI1 phosphatase: How to validate true interactions?
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Methodology:
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Perform bimolecular fluorescence complementation (BiFC) with SLAC1-YFPᶜ and ABI1-YFPᴺ in guard cell protoplasts .
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Quantify interaction strength using fluorescence lifetime imaging microscopy (FLIM) to detect FRET efficiency changes .
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Correlate with anion current measurements: Functional ABI1-SLAC1 binding should reduce OST1-activated currents by ≥70% .
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Structural and Functional Optimization
What criteria define high-quality SLAC1 antibodies for cryo-EM studies?
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Resolution: Antibody-antigen complex should resolve ≤3.5 Å to visualize latch/plug domain rearrangements .
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Epitope stability: Epitopes must withstand grid freezing conditions (e.g., 0.1% glutaraldehyde for 30 s).
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Validation metric: Antibody should recover ≥90% of SLAC1 channels in single-particle analysis from native membranes .
