ELISA for IgM or IgG

ELISA for IgM or IgG

• • Precoating plate with Ab (RAM Zymed 0,5mg/ml diluited 1:500 or GAM Zymed 2mg/ml 1:10.000) in PBS, o/n 4°C, 100μl /well

  • Wash 3 times with PBS 200 μl/well

  • Blocking di 3% milk or 3% FCS or 3% BSA in PBS/0.05%Tween 200μl/well minimum 30min RT

  • Wash 3 times with PBS 200 μl/well

  • Add samples (mouse serum 1:5.000 or supernatant (1:1) or 30.000 cell lysate in 100μl PBS/well) e incubate 1h RT or O/N at 4°C

  • Wash 3 times with PBS/Tween 0.05% 200 μl/well

  • Add 100 μl anti mu chain HRP-Ab (Zymed) 1:2000 or anti IgG-HRP-Ab (Zymed) 1:10.000 in PBS 45 min RT

  • Wash 3 times with PBS/Tween 0.05% 200 μl/well

  • Add 100 μl/well supersensitive for ELISA (Sigma T4444)

  • Block with 1M Hcl (100 μl/well)

  • Measure at 450nm