FUT7 Antibody

Basic Research Questions

How to validate FUT7 antibody specificity in experimental models?

• Perform Western blot analysis using lysates from FUT7-knockdown or overexpression cell lines (e.g., Jurkat T-cells , MHCC97 hepatocarcinoma cells , or A549 lung adenocarcinoma cells ).

• Expected band size: ~39–50 kDa (aligns with protein molecular weight predictions ).

• Include controls such as:

  • siRNA-transfected cells (e.g., FUT7 siRNA reduces protein levels by >50% in MHCC97 cells ).

  • Non-targeting siRNA (siNC) to confirm specificity .

• Pair with functional assays (e.g., adhesion/invasion assays) to correlate antibody detection with biological outcomes .

What experimental models are optimal for studying FUT7 in cancer biology?

How to quantify FUT7 in clinical samples?

• Immunohistochemistry (IHC): Use intensity scoring (0–3 scale) in bladder cancer tissues vs. normal controls .

• ELISA: Detect FUT7 in serum (AUC = 0.754 for bladder cancer diagnosis ).

• Flow cytometry: Measure SLe^X levels (e.g., reduced fluorescence intensity after FUT7 knockdown ).

Advanced Research Questions

How to resolve contradictory data on FUT7’s role in proliferation vs. apoptosis?

• Context-dependent analysis:

  • Pro-proliferative effects: Observed in A549 (lung) and Jurkat (ALL) cells via EGFR/Akt and adhesion pathways .

  • Pro-apoptotic effects: FUT7 knockdown in Jurkat cells increases apoptosis by 35% .

• Methodological considerations:

  • Use isogenic cell lines (knockdown vs. overexpression) to isolate FUT7-specific effects .

  • Combine transcriptomics (e.g., RNA-seq) with phospho-proteomics to identify downstream signaling nodes .

How to integrate FUT7 antibody data with immune microenvironment profiling?

• Correlative approaches:

  • Link FUT7 IHC scores with CD8+/CD4+ T-cell infiltration levels in bladder cancer (Spearman r = 0.42–0.58 ).

  • Use multiplex IF to co-stain FUT7 and immune checkpoints (e.g., PD-L1) in tumor sections .

• Functional assays:

  • Co-culture FUT7-overexpressing cancer cells with peripheral blood mononuclear cells (PBMCs) to assess T-cell activation .

What strategies address variability in FUT7 glycosylation across models?

• Glycan-specific antibodies: Use anti-SLe^X antibodies (e.g., KM93 ) to confirm functional FUT7 activity.

• Enzymatic deglycosylation: Treat lysates with PNGase F to distinguish core protein vs. glycosylated forms .

• Orthogonal validation: Pair Western blot with lectin arrays (e.g., E-selectin-Fc binding assays ).

Methodological Best Practices

• Antibody validation: Always include recombinant FUT7 protein as a positive control .

• Data reproducibility: Replicate findings across ≥3 cell models (e.g., leukemia, carcinoma, lymphoma ).

• Clinical correlation: Use TCGA or PrognoScan databases to assess FUT7’s prognostic value (e.g., hazard ratio = 1.8 in bladder cancer ).