WAKL3 Antibody

Here’s a structured collection of FAQs for researchers working with WAKL3 antibodies, designed to address academic research scenarios with methodological rigor and data-driven insights:

Advanced Research Questions

• How to resolve contradictions in WAKL3 localization studies?

  • Conflict: Some studies report plasma membrane localization, while others emphasize cell wall association.

  • Resolution: Use differential extraction protocols (e.g., SDS/DTT for cell wall-bound proteins vs. Triton X-100 for membrane proteins). WAKL6, a homolog, was exclusively found in insoluble fractions, confirming cell wall linkage .

  • Method: Immunolocalization with confocal microscopy using chlorophyll-autofluorescence quenching for clearer signal detection in mesophyll cells .

• What mechanisms explain WAKL3’s role in pathogen defense?

  • Hypothesis: WAKL3 may bind pathogen-associated molecular patterns (PAMPs) via its extracellular domain, analogous to WAK1’s interaction with pectin during fungal infection .

  • Experimental design:

    • Perform co-immunoprecipitation (Co-IP) with fungal β-glucan or chitin fragments.

    • Compare transcriptional responses of WAKL3 KO vs. wild-type plants under Pseudomonas syringae infection.

• How does WAKL3 antibody performance vary across developmental stages?

  • Data: Promoter-GUS assays show WAKL3 expression peaks during root elongation and flower maturation.

  • Validation strategy: Time-course WB/IF experiments using synchronized Arabidopsis populations. For example, WAKL3 signal intensifies in root tips at 14 days post-germination .

Methodological Best Practices

• Antibody validation:

  • Use KO lines as negative controls for WB/IF .

  • For IF, optimize fixation (e.g., 4% formaldehyde) to preserve cell wall integrity .

• Data interpretation:

  • Account for truncated isoforms (e.g., secreted WAKL7) that may cross-react with WAKL3 antibodies .

  • Use RRID portals (e.g., AntibodyRegistry.org) to track validation data and avoid discontinued reagents .

Key Challenges in WAKL3 Research