Hrp conjugated rabbit anti mouse igs antibody

Approximately 80 mg of lyophilized Muc5ac purified from either wild-type or Fut2-null mice were resuspended in 100 μL of MQ-water. Serial dilutions were prepared (800; 80; 40; 20; 10 and 5 μg/μL) and 2 μL from each solution were sequentially applied as separate dots onto nitrocellulose membranes (Amersham Hybond-ECL Membrane, GE Healthcare, Buckinghamshire, UK). Membranes were blocked for 1 hr with PBS containing 5% BSA prior to incubation over-night at 4 °C with BG6, SPM279 and PM7 diluted in PBS with 1% BSA, as described in Table 1. As a negative control, primary antibody was replaced by PBS with 1% BSA. Blots were washed three times with PBS-Tween 0.01% and incubated with HRP-conjugated rabbit anti-mouse Igs antibody (DAKO, Glostrup, Denmark), diluted 1:1000 in PBS containing 1% BSA, for 1 hr at RT. Membranes were washed as described above and developed using ECL (Amersham ECL Western Blotting detection reagents, GE Healthcare, Buckinghamshire, UK).