Costar 9018 elisa plates

Sera were collected from the immunized animals to monitor the antibody response by ELISA. Corning Costar 9018 ELISA plates (Corning Costar, Inc., Corning, NY, USA) were coated with Ag85B (5 µg/ml) or ESAT-6 (5 µg/ml). The plates were blocked with PBS containing 1% bovine serum albumin (BSA) (Bovogen Biologicals PTY., Ltd., VIC, Australia). Sera were added at serial 2-fold dilution (beginning at a 1/100 dilution). After washing, and adding horseradish peroxidase-conjugated goat anti-mouse IgG, IgG1 and IgG2c (SouthernBiotech, Birmingham, AL, USA) were diluted at 1/10,000, 1/1,000 and 1/1,000 separately in blocking buffer (PBS containing 1% BSA). Plates displayed color by o-phenylenediamine substrate. Antibody titers were expressed as reciprocal end point titers.

Sera were collected from the immunized mice to monitor the antibody response by ELISA. Corning Costar 9018 ELISA plates (Corning Costar, Inc., Corning, NY, USA) were coated with Ag85B (5 µg/ml) overnight at 4°C. The plates were blocked with PBS containing 1% bovine serum albumin (BSA; Bovogen Biologicals Pty Ltd., East Keilor, VIC, Australia). Sera were added at serial 2-fold dilution (beginning at a 1/50 dilution). After washing, and addition of horseradish peroxidase conjugated goat anti-mouse IgG, IgG1 and IgG2c (SouthernBiotech, Birmingham, AL, USA) diluted at 1/5,000, 1/1,000 and 1/1,000, respectively in blocking buffer (PBS containing 1% BSA) for 1 h, the plates displayed the staining by o-phenylenediamine substrate. Antibody titers were expressed as reciprocal end point titers. The mean antibody titers were the antilog of the average logarithm for antibody titer of 5 mice.