Western Blot protocol specific for PAPP A Antibody (NB200-401)

1) Load Native human htPAPP-A (PAPP-A/proMBP complex) onto gradient 3,5 ? 7,5 % gel for SDS-PAGE.
2) Electrotransfer 60 min at 110 V,
3) Blocking 5% milk/PBST 1 hour at RT
4) NB200-401 10 ug/ml in PBST, incubation 1 hour at RT,
5) Secondary antibody rabbit anti-mouse IgG-HRP-linked 1:1000 in 5% milk/PBST, incubation 1 hour at RT.
6) Diaminobenzidine as a substrate