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Complement Assay

Assess complement activity/lysis
🎯 Difficulty ★★☆ Medium Duration About 3–4 h 🎯 Use Evaluate antibody-dependent complement lysis and complement pathway activation
📖 Principle
Antibody-target binding activates the classical complement pathway; membrane attack complex lyses cells, measured as cytotoxicity
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📋 Workflow

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1

Target cell preparation

1 h
Culture and label target cells, count and resuspend at suitable density
💡 Tip Load dye/radio label to detect lysis
⚠️ Caution High viability ensures reliable results
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2

Antibody sensitization

30 min
Incubate targets with excess antibody to sensitize them for complement activation
💡 Tip Excess antibody ensures saturation
⚠️ Caution Wash off free antibody to control background
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3

Complement treatment

1 h
Add complement source (human/rabbit serum) and incubate to activate lysis
💡 Tip Control complement titer/stability
⚠️ Caution Keep complement cold to prevent inactivation
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4

Lysis detection

1 h
Measure released marker in supernatant or stain live cells to calculate lysis percentage
💡 Tip Include spontaneous and maximum release controls
⚠️ Caution High background masks differences
🧪 Materials for this step (click to shop by spec/brand)

⚠️ 5 Common Beginner Mistakes

  • Inactivated complement gives false negatives
  • High spontaneous release masks complement activity
  • Insufficient antibody reduces lysis
  • Excess complement causes non-specific lysis
  • Missing maximum release hampers lysis calculation

❓ FAQ

+Which controls are necessary?
Spontaneous (no complement), maximum (lysis agent) and no-antibody controls are essential.
+How to improve complement lysis?
Optimize antibody density, complement titer, target sensitivity and matched source.
+How to store active serum?
Use fresh or aliquot at -80°C, avoiding repeated freeze-thaw and heat inactivation.

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Data authenticity: Product categories from JW Supply Chain standard taxonomy; method→step→product connected. Last updated: 2026-08-23 · v3.0