FBL16 Antibody

Here’s a structured collection of FAQs for FBL16 (FBXL16) antibodies tailored to academic research, incorporating technical depth, methodological guidance, and data from diverse sources:

Advanced Research Questions

How to resolve contradictions in FBL16 localization data across studies?

• Root Causes:

  • Antibody cross-reactivity with homologous F-box proteins (e.g., FBXW7) .

  • Tissue fixation differences (e.g., paraffin-embedded vs. fresh-frozen) .

• Solutions:

  • Validate with orthogonal methods (e.g., RNAscope® for mRNA vs. antibody staining) .

  • Compare multiple clones (e.g., polyclonal vs. monoclonal) .

How to design experiments investigating FBL16’s role in E3 ubiquitin ligase complexes?

• Functional Assays:

  • Co-immunoprecipitation (Co-IP): Use anti-SKP1/CUL1 antibodies to confirm SCF complex interaction .

  • Ubiquitination Assay: Express HA-tagged ubiquitin + FBL16 in HEK293T cells; detect shifts in substrate molecular weight .

• Critical Controls:

  • Dominant-negative FBL16 (F-box domain deletion) .

  • Proteasome inhibition (MG132) to stabilize ubiquitinated substrates .

What are the limitations of current FBL16 antibodies in mechanistic studies?

• Key Issues:

  • Lack of isoform-specific reagents (e.g., spinal cord injury-associated Scirr1 variant) .

  • Limited cross-species reactivity (validated only in human) .

• Workarounds:

  • CRISPR/Cas9 knock-in epitope tags (e.g., FLAG) for endogenous tracking .

  • Combine antibody data with transcriptomics (e.g., RNA-seq of FBL16-silenced cells) .

Observed vs. Predicted Molecular Weight

Methodological Notes

• Epitope Mapping: Abcam’s antibody targets residues in the synthetic peptide ILNGLFWYFSACEKCVLAQVCKA , while Proteintech’s uses a fusion protein .

• Ethical Compliance: All products are research-use-only; licensing required for diagnostic/therapeutic applications .