Ohne Titel

Immunoblotting

SDS-PAGE was prepared using antigen lysate and NuPAGE MOPS protein ladder in Novex gel

SDS-PAGE was ran at 200V to separate the lysate constituents

Antigens transferred onto nitrocellulose paper. Blocking solution then anti-sera were added. Alkaline phosphatase-conjugated reveal antibody was then added

Blot was developed with BCIP/NBT and images were acquired

Static adhesion assay

ICAM 1 in coating buffer added and incubated overnight. Blocking buffer was then added

Anti-DBL sera and infected erythrocytes incubated in tubes together to allow for antibody opsonisation.

Blocking buffer discarded from the plate and tube contents added to appropriate wells. Infected erythrocyte standard curve wells were also constructed

Plate images acquired and absorbance read at 405nm