Rabbit anti c myc antibody

For analysis of piliation on strains grown on solid medium, immunoelectron microscopy was performed as described previously (52 (link)). Briefly, Formvar/carbon-coated copper grids were used to lift cells directly from 18-h-old colonies and fixed for 15 min by addition of a drop (17 μl) of 0.2% glutaraldehyde and 4% paraformaldehyde in Dulbecco’s PBS (DPBS; Fisher) onto the grids. The grids were washed 3 times with 1% bovine serum albumin (BSA; Sigma) in DPBS and blocked in 0.1% gelatin (Aurion, Inc.) in DPBS for 30 min. The grids were washed once with 1% BSA in DPBS and incubated with a 1:10 dilution of rabbit anti-c-Myc antibody (Sigma) for 1 h. The grids were washed three times with 1% BSA in DPBS and incubated with 0.1% gelatin in PBS for 30 min. They were then washed once with BSA in DPBS and incubated with goat anti-rabbit IgG antibody conjugated to 12-nm gold particles (1:20 dilution; Jackson ImmunoResearch Laboratories) for 1 h. The grids were washed five times in water for 3 min each and then negatively stained with 1% uranyl acetate for 1 min. All washes and incubations were with 17 μl and performed at room temperature. The liquid on the grids after each step was carefully wicked away using a Whatman paper. Grids were viewed using an FEI Tecnai Spirit G2 transmission electron microscope.