Goat ant rabbit igg ir700

B. burgdorferi strains B31A3 and A3HtrAOE were harvested from BSK II medium and washed two times with DPBS. A total of 2 × 10⁷ cells from each were separated by SDS-PAGE and analyzed by western blot using rabbit polyclonal anti-p66, rabbit anti-HtrA and mouse monoclonal anti-FlaB. Secondary antibodies used were goat ant-rabbit IgG IR700 and goat anti-mouse IgG IR800 (Rockland Immunochemicals). Digital images of western blot bands were analyzed using a Bio Rad GD/Chemi Documentation System and Quantity One Quantitation Software (Bio Rad) to determine optical densities. Each band was quantitated three times (left end, middle, right end) and the combined data from two independent experiments was tested for statistical significance by unpaired t test (Graph Pad, La Jolla, CA).

The procedure for identification of detergent-resistant membranes (DRM) has been previously described (LaRocca et al., 2010 (link), Toledo et al., 2014 ). Briefly, B31 spirochetes were treated with 1% Triton X-100 for 16 hours at 4°C. The lysate was separated on a discontinuous OptiPrep density gradient and the fractions were analyzed for α-asialo-GM1, p66 and HtrA by slot blot using rabbit polyclonal antibodies to ganglioside asialo GM1, p66 and HtrA, and mouse anti-DnaK. Secondary antibodies used were goat ant-rabbit IgG IR700 and goat anti-mouse IgG IR800 (Rockland Immunochemicals).