Immune electrophoresis

 

 

Basis:                       Combination of electrophoresis and immune precipitation

Gel:                           Natives agarose gel

                                 (no acrylamide gel, no SDS !)

Buffer:                      barbital buffer (18g/l) pH = 8.4 + 1g sodium azide /l

Agar:                        1-1.5 % in buffer

Elpho:                       6 V/cm, about 45 min

Immune diffusion:     6h/37°C or 24h/4°C in wet chamber

Stain:                       

·       put gel in water to remove soluble protein!!

·       dry gel

·       stain: 0.05% Coomassie Brilliant Blue G250

·       in acetic acid / ethanol / water 1:5:4 (v/v/v), 5-10 min

·       destain background with acetic acid / ethanol / water 1:5:4 (v/v/v)