clec-91 Antibody

CLEC9A (C-type lectin domain family 9 member A) is an activation receptor involved in dendritic cell function and immune response regulation. Below are research-focused FAQs addressing key experimental considerations for working with CLEC9A antibodies, organized by complexity level:

Advanced Research Questions

How to resolve discrepancies in CLEC9A signaling studies across publications?

Common sources of variability and solutions:

What functional assays confirm CLEC9A's role as an activation receptor?

• Cytokine profiling: Measure TNF-α/IL-12p70 via ELISA after antibody cross-linking

• Phagocytosis assay: Compare zymosan uptake in CLEC9A+ vs. CLEC9A- dendritic cells

• Calcium flux: Use Fluo-4 AM staining with receptor ligation

How to optimize CLEC9A detection in low-abundance primary cells?

• Pre-enrichment strategy:

  • Magnetic sorting with CD303 microbeads

  • High-sensitivity detection: Phycoerythrin tandem dyes > APC

• Amplification: Tyramide signal amplification (TSA) with 0.1% Triton X-100 permeabilization

Technical Challenge Solutions

Addressing nonspecific binding in CLEC9A immunohistochemistry

• Blocking cocktail: 5% mouse serum + 50 μg/ml human IgG

• Validation controls:

  • Tissue from CLEC9A-/- mice (if available)

  • RNAscope® ISH correlation

Standardizing cross-linking conditions for CLEC9A activation studies

Data Interpretation Guidelines

Distinguishing CLEC9A isoforms in Western blots

Quantitative analysis challenges in CLEC9A dimerization studies

• Use non-reducing PAGE + Förster resonance energy transfer (FRET)

• Reference data: CLEC9A forms stable dimers with Kd = 1.8 × 10⁻⁷ M