HSD2 Antibody

Here’s a structured FAQ collection for HSD2 antibody research, derived from peer-reviewed studies and technical documentation:

Advanced Research Questions

How to resolve contradictory reports of HSD2 localization in vascular vs. epithelial cells?

• Experimental design considerations:

  Variable	Impact on Results
  Tissue processing	Postmortem delay reduces vascular staining
  Antibody clone	HUH23 vs. MAB8630 show endothelial vs. epithelial preference

  • Solution: Combine multiple clones (e.g., HUH23 + MAB8630) for dual-labeling experiments .

What mechanisms underlie HSD2’s role in glucocorticoid signaling beyond enzymatic activity?

• Key findings:

  • SUMOylation at K266 modulates cortisol-induced MR nuclear translocation .

  • Dimerization (80 kDa) and trimerization (120 kDa) observed in CEM-C7 cells .

• Method: Co-immunoprecipitation with anti-SUMO1 antibodies + proximity ligation assays .

Methodological Challenges

How to optimize HSD2 detection in low-abundance tissues?

• Signal amplification:

  • Tyramide-based systems for IHC

  • Pre-adsorption with recombinant HSD11B2 protein (10 µg/mL) reduces background

• Quantification: Use renal cortical tubules as internal intensity reference .

Why do molecular weights vary between studies?

• Critical variables:

  Factor	Effect
  Glycosylation	+3–5 kDa shift
  Reducing conditions	Eliminates 48 kDa renal isoform

  • Always include molecular weight markers spanning 35–50 kDa .

Cross-Disciplinary Applications

Can HSD2 antibodies be used in neuroendocrine studies?

• Evidence:

  • Phox2b co-expression in NTS neurons enables dual-labeling for aldosterone signaling studies .

  • Protocol: Sequential IHC (HSD2 → Phox2b) with heat-mediated antibody stripping .

Troubleshooting Table