📋 Workflow
Full workflow, products add-to-cart directly1
Coat capture antibody
OvernightCoat ELISpot plate with cytokine capture antibody, incubate overnight at 4°C then block
💡 Tip Even coating ensures spot formation
⚠️ Caution Wash gently to avoid antibody loss
🧪 Materials for this step (click to shop by spec/brand)
2
Cell seeding and stimulation
24–48 hAdd PBMCs and stimulate with antigen/peptide pool, incubate to induce cytokine secretion
💡 Tip Include no-antigen negative control
⚠️ Caution Optimal density avoids spot merging
🧪 Materials for this step (click to shop by spec/brand)
3
Development
2 hWash, add detection antibody and enzyme substrate, develop spots, stop and air-dry
💡 Tip Control development time
⚠️ Caution Overcrowded spots uncountable
🧪 Materials for this step (click to shop by spec/brand)
4
Spot counting and analysis
30 minCount spots per well with ELISpot reader, compute frequency of antigen-specific cells
💡 Tip Consistent counting parameters
⚠️ Caution Exclude non-specific background spots
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Uneven coating distorts spots
- ② Too many cells merge spots
- ③ Over-development masks individual spots
- ④ Missing negative control for background
- ⑤ Harsh washing loses cells/antibody
❓ FAQ
+ELISpot vs ELISA?
ELISpot measures secreting cell frequency/spots; ELISA measures total secreted amount.
+How to judge positive spots?
Positive spots clearly exceed background in size/density with regular shape.
+How to set cell density?
Depends on frequency; use cell titration to stay in countable range.