Y18D10A.9 Antibody

Y18D10A.9 Antibody: Frequently Asked Questions for Academic Researchers

This FAQ addresses critical methodological and conceptual challenges in working with Y18D10A.9 antibodies, focusing on experimental design, data interpretation, and advanced applications in C. elegans research. Content is derived from peer-reviewed protocols, genetic interaction studies, and functional characterization data .

Advanced Research Questions

How to design CRISPR-based experiments to study Y18D10A.9 genetic interactions?

• Workflow:

  • Design crRNA targeting Y18D10A.9 using the 20-nucleotide sequence upstream of the NGG PAM motif .

  • Co-inject Cas9 ribonucleoprotein complexes with homology-directed repair (HDR) templates (Figure 2.6-2.9) .

  • Screen F2 progeny for homozygous mutants using PCR and Sanger sequencing.

Genetic interaction analysis:

How to resolve contradictions in Y18D10A.9 functional data across studies?

• Analysis framework:

  • Compare strain-specific genetic backgrounds (e.g., N2 vs. Hawaiian variants).

  • Control for temperature sensitivity (e.g., 20°C vs. 25°C culture conditions) .

  • Quantify protein localization dynamics using time-lapse imaging of GFP-tagged strains (Protocol 2.4) .

Case example: Discrepancies in sperm activation phenotypes may arise from differences in:

• Spermatogenesis staging (Figure 4.35-4.37) .

• Antibody penetration efficiency in dissected gonads vs. intact tissues.

What advanced techniques are required to study Y18D10A.9 in signaling pathways?

• Integrated methodology:

  • Co-immunoprecipitation: Use endogenously tagged FLAG::Y18D10A.9 strains (Figure 4.31-4.33) .

  • Live imaging: Combine mCherry-histone and GFP-tubulin markers to track mitotic defects (Figure 4.36) .

  • Phosphoproteomics: Perform LC-MS/MS on immunoprecipitated complexes using TiO2 enrichment.

Technical Notes

• Antibody storage: Aliquot in stabilizing buffer (50% glycerol, 0.1% BSA) at -80°C for long-term preservation.

• Quantitative analysis: Use Fiji/ImageJ macros for automated signal quantification in sperm activation assays .