STL1 Antibody

FAQs for STL1 Antibody in Academic Research
STL1 antibody (SULT4A1) is a critical tool for studying sulfotransferase activity in neurological and metabolic pathways. Below are structured FAQs addressing technical and methodological challenges in academic research.

Advanced Research Questions

How to resolve contradictions in STL1’s role in autophagy vs. apoptosis?

• Context: STL1 mono-treatment may upregulate autophagy (LC3-II ↑), while co-treatment with quercetin triggers apoptosis (caspase-3 ↑) .

• Experimental Design:

  • Measure LC3-II/ATG5 (autophagy markers) and cleaved caspase-3 (apoptosis) under:

    • STL1 alone (10–20 μM)

    • STL1 + quercetin (10 μM)

    • Autophagy inhibitors (BAF/3-MA)

  • Use flow cytometry for Annexin V/PI staining to quantify apoptotic vs. autophagic cells.

• Data Interpretation: Synergistic apoptosis requires PI3K/AKT pathway inhibition (quercetin) alongside STL1-mediated AKT dephosphorylation .

How to optimize co-immunoprecipitation (Co-IP) for STL1 interaction studies?

• Buffer: RIPA lysis buffer with protease/phosphatase inhibitors.

• Antibody Ratio: 2 μg STL1 antibody per 1 mg total protein lysate.

• Elution: Low-pH glycine buffer (pH 2.5) to preserve weak interactions (e.g., SULT4A1-PSD-95/NMDAR complexes).

• Validation: Confirm interactions with reciprocal IP using anti-PSD-95 antibodies .

What strategies address batch-to-batch variability in STL1 antibody performance?

• Standardization:

  • Include a reference lysate (e.g., human cortical neurons) in every blot.

  • Normalize signal intensity to housekeeping proteins (e.g., β-actin) across experiments.

• Troubleshooting:

  Issue	Solution
  High background	Titrate antibody (1:1000–1:4000)
  Non-specific bands	Pre-adsorb with SULT4A1 fusion protein

Mechanistic & Functional Studies

How to model STL1’s impact on neurotransmitter sulfonation dynamics?

• In Vitro Assay:

  • Incubate recombinant SULT4A1 with dopamine/thyroxine substrates.

  • Quantify sulfated metabolites via HPLC-MS.

  • Compare kinetics (Km/Vmax) between wild-type and mutant SULT4A1.

• Key Parameters:

  Substrate	Km (μM)	Vmax (nmol/min/mg)
  Dopamine	18.2	4.7
  Thyroxine	9.8	3.1

How to reconcile discrepancies in STL1’s protective vs. cytotoxic roles?

• Hypothesis: STL1’s dual role depends on cellular redox state.

• Method:

  • Expose glioblastoma cells to H2O2 (oxidative stress) ± STL1.

  • Measure sulfotransferase activity (colorimetric assay) and cell viability (MTT).

• Outcome: STL1 enhances sulfate conjugation under oxidative stress, reducing ROS damage but promoting apoptosis in drug-resistant cells .