mouse anti rainbow trout antibodies  (Bio-Rad)

Journal: Vaccines

Article Title: Vaccination with Ectoparasite Proteins Involved in Midgut Function and Blood Digestion Reduces Salmon Louse Infestations

doi: 10.3390/vaccines8010032

Figure Lengend Snippet: Antibody response in vaccinated fish. ( A ) The recombinant proteins were produced in E. coli and analyzed by polyacrylamide gel electrophoresis (PAGE) and Western blot (WB). Ten micrograms of recombinant proteins was loaded onto a 12% SDS-polyacrylamide gel and stained with Coomassie Brilliant Blue or transferred to a nitrocellulose membrane. For Western blot analysis, pooled sera collected from vaccinated salmons before salmon louse infestation at approximately week 10 post-initial vaccination were used as primary antibodies. The membrane was then incubated with mouse anti-rainbow trout antibodies and revealed with a goat anti-mouse IgG-HPR conjugate. ( B ) Antibody titers were determined in vaccinated and control fish by ELISA. Values of D.D. at 450 nm in vaccinated and control groups were compared by Student’s t -test with Welch’s correction for unequal variances ( p = 0.05).

Article Snippet: Plates were washed three times with PBS/0.05% Tween 20, and 100 μL/well of mouse anti-rainbow trout antibodies (Bio-Rad Laboratories, Inc.), diluted (1:1000, v / v ) in blocking solution, was added and incubated for 1 h at RT.

Techniques: Recombinant, Produced, Polyacrylamide Gel Electrophoresis, Western Blot, Staining, Membrane, Incubation, Control, Enzyme-linked Immunosorbent Assay