C02C2.6 Antibody

FAQs for C02C2.6 Antibody in Academic Research

Advanced Research Questions

• How to resolve contradictions in C02C2.6 interaction data across studies?

  • Analysis framework:

    • Compare bait protein tags (e.g., GST vs. FLAG) and lysis buffer stringency (e.g., RIPA vs. mild detergents) .

    • Use orthogonal methods: Co-IP followed by HMS-PCI mass spectrometry vs. yeast two-hybrid screens .

    • Example conflict: C02C2.6 binding to HIF-1 observed under hypoxia but not normoxia. Replicate conditions with O₂ sensors and validate via targeted MS (PRM/SRM) .

• What strategies identify C02C2.6’s post-translational modifications (PTMs) in aging models?

  • Workflow:

    1. Enrich C02C2.6 via IP under denaturing conditions (6 M urea).

    2. Digest with trypsin/Lys-C and analyze via LC-MS/MS with phosphopeptide enrichment (TiO₂) or ubiquitin remnant motif (K-ε-GG) antibodies .

    3. Cross-reference with C. elegans aging datasets (e.g., GFAT-1-linked proteostasis decline) .

• How to design a genetic interaction screen using C02C2.6 Antibody?

  • Approach:

    • Combine RNAi (e.g., hif-1, egl-9) with C02C2.6 IF in let-60(gf) RAS/MAPK hyperactivation mutants .

    • Quantify vulval development defects (e.g., multivulva phenotype) and correlate with C02C2.6 subcellular localization shifts (nuclear vs. cytoplasmic) .

Methodological Best Practices

• Cross-reactivity checks: Test antibody against C. briggsae homologs (e.g., CB4856 strain) to assess species specificity .

• Quantitative analysis: Use Fiji/ImageJ macros for IF intensity normalization to housekeeping proteins (e.g., ACT-1) .

• Data rigor: Report antibody lot numbers, fixation times, and protease inhibitor cocktails used (e.g., PMSF vs. EDTA-free formulations) .