PEPR1 Antibody
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- This study presents the crystal structure of the extracellular LRR domain of PEPR1 (PEPR1LRR) in complex with AtPep1, a 23-amino acid endogenous peptide. PMID: 25475059
- AtPeps signal through interaction with PepR1, a plasma membrane protein that can generate cGMP. In a signaling cascade, the cGMP-activated channel CNGC2 participates in AtPep- and PepR1-dependent inward Ca2+ conductance and the resulting cytosolic Ca2+ elevation. PMID: 21088220
- Gene At1g73080 encodes a leucine-rich repeat receptor kinase called PEPR1, which serves as a receptor for the AtPep1 peptide. [PEPR1] PMID: 16785433
Q&A
FAQs for PEPR1 Antibody in Academic Research
What experimental designs leverage PEPR1 antibodies to study receptor-ligand interactions?
Advanced Applications:
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Co-immunoprecipitation (Co-IP): Use PEPR1 antibodies to pull down complexes with BIK1 or BAK1 in Arabidopsis lysates pretreated with Pep1. Validate interactions via kinase assays .
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Ligand-induced endocytosis: Track PEPR1 internalization in live cells using fluorescently labeled antibodies alongside TAMRA-Pep1, with serk/bak1 mutants as controls .
| Assay | Key Controls | Critical Parameters |
|---|---|---|
| Co-IP | bik1 mutants, untreated lysates | Protease inhibitors, crosslinkers (e.g., DSS) |
| Endocytosis | bak1-3serk4-1 mutants | Time-lapse imaging (40-min chase) |
How to resolve contradictions in PEPR1 phosphorylation data across studies?
Troubleshooting Approach:
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Kinase activity dependency: Compare phosphorylation in kinase-dead PEPR1 variants versus wild type using phospho-specific antibodies .
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Context-specific signaling: Test immune triggers (e.g., ethylene, Botrytis cinerea) to differentiate basal vs. induced phosphorylation states .
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Buffer optimization: Include phosphatase inhibitors (e.g., NaF) and reduce SDS concentrations to preserve post-translational modifications .
What methods optimize PEPR1 antibody performance in challenging tissues (e.g., roots)?
Protocol Recommendations:
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Tissue-specific extraction: Use Tris-HCl buffers with 1% Triton X-100 for root membrane proteins .
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Crosslinking: Apply formaldehyde fixation to stabilize PEPR1-BAK1 complexes before homogenization .
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Signal amplification: Pair antibodies with tyramide-based amplification for low-abundance PEPR1 detection in immunolocalization .
How to design controls for PEPR1 antibody use in pathogen interaction studies?
Advanced Controls:
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Pathogen-free vs. infected: Compare PEPR1 levels in Pseudomonas syringae-infected vs. mock-treated plants .
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Effector protein impact: Include Pst DC3000 strains lacking T3Es (e.g., ΔavrPto) to assess effector-specific PEPR1 regulation .
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Dual localization: Co-stain with endosomal markers (e.g., Rab5) to confirm antibody specificity in internalization assays .
How to validate PEPR1 antibody compatibility with quantitative techniques (e.g., ELISA)?
Validation Workflow:
