Selected article for: "serum dilution and western blot"
Author: Tucci, Paula; Estevez, Verónica; Becco, Lorena; Cabrera-Cabrera, Florencia; Grotiuz, Germán; Reolon, Eduardo; Marín, Mónica
Title: Identification of Leukotoxin and other vaccine candidate proteins in a Mannheimia haemolytica commercial antigen
  • Document date: 2016_9_19
    • ID: 71omypkg_18
    Snippet: Both rabbits were immunized subcutaneously with 50 ug of recombinant LKT, on days 0 and 21 of the immunization protocol. Blood samples were taken on days 21, 45 and 115, and sera were prepared and stored at −20°C. Serum titers were determined by ELISA using 0.1 μg/well of recombinant LKT for coating, serum dilutions of 1:100 to 1:100.000, and conjugate dilution of 1:10.000 (Goat antirabbit IgG (whole molecule)-Peroxidase, Sigma). High titer s.....
    Document: Both rabbits were immunized subcutaneously with 50 ug of recombinant LKT, on days 0 and 21 of the immunization protocol. Blood samples were taken on days 21, 45 and 115, and sera were prepared and stored at −20°C. Serum titers were determined by ELISA using 0.1 μg/well of recombinant LKT for coating, serum dilutions of 1:100 to 1:100.000, and conjugate dilution of 1:10.000 (Goat antirabbit IgG (whole molecule)-Peroxidase, Sigma). High titer sera were pooled and purified by protein A chromatography (HiTraprProtein A, GE Healthcare). Purified IgG reactivity against native and recombinant LKT was evaluated by Western blot as described below.

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