SEC31A Antibody

FAQs for SEC31A Antibody in Academic Research

Advanced Research Questions

• How to resolve discrepancies in SEC31A molecular weight across studies?

  • Observed variations: SEC31A migrates at 118–134 kDa due to:

    • Isoforms: Alternative splicing generates ~133 kDa (full-length) and truncated variants .

    • Phosphorylation: COPII assembly regulation modifies electrophoretic mobility .

  • Strategies:

    • Use deglycosylation enzymes (e.g., PNGase F) to assess glycosylation impact .

    • Compare migration in multiple cell lines (HeLa vs. HepG2) .

• What mechanisms link SEC31A gene aberrations to inflammatory myofibroblastic tumors (IMTs)?

  • Chromosomal rearrangements: SEC31A-ALK fusions (e.g., SEC31A exon 20–ALK exon 20) drive constitutive kinase activity .

  • Validation workflow:

    • FISH or RNA-seq to detect fusion transcripts .

    • ALK inhibitor sensitivity assays (e.g., crizotinib) in SEC31A-ALK+ IMT models .

• How to differentiate SEC31A isoforms in subcellular trafficking studies?

  • Isoform-specific antibodies: Most commercial antibodies target the N-terminal WD40 domain (common to all isoforms) . For isoform resolution:

    • Design custom antibodies against unique C-terminal epitopes .

    • Combine siRNA targeting specific exons with endogenous SEC31A detection .

  • Functional readouts:

    Isoform	Trafficking Role	Assay
    Full-length	ER export of bulky cargo (e.g., collagen)	FRAP-based ERES dynamics
    Truncated	Regulatory (dominant-negative)	COPII assembly inhibition assays

Methodological Notes

• Antibody cross-reactivity: While SEC31A antibodies are cited for human, mouse, and rat , always validate cross-species reactivity using tissue lysates (e.g., zebrafish SEC31A shares ~60% homology with human) .

• Data contradictions: If SEC31A signal persists in knockout cells, check for off-target binding to SEC31B (40% homology) . Use tandem mass spectrometry to confirm immunoprecipitated proteins .