Goat anti rabbit igg texas red secondary antibody

PMN were sorted using anti-Ly6C/Brilliant Violet 421 and anti-Ly6G/PerCpCy5.5 antibodies (Biolegend) and then incubated in RP-10(−) medium with PGE₂ (final concentration 1 μM) or vehicle (ethanol) for 60 min at 37°C in 7% CO₂. L. monocytogenes SD2710 (GFP+) was added and the plate was centrifuged for 5 min at 300 x g followed by incubation for 5 min at 37°C in 7% CO₂. Polyclonal rabbit anti-Listeria O primary antisera (Difco) and goat anti-rabbit IgG–Texas Red secondary antibody (Thermo Fisher) were used to perform differential “in/out” staining as described previously (27 (link)). Cells were visualized using a Zeiss Axio Imager.Z1 with a 100x/1.4 NA PlanApo oil immersion objective and analyzed with AxioVision software. Slides were blinded and examined by two different investigators; average values are reported.

Antibodies recognizing p-ERK1/2 (T202/Y204) (#4370), ERK1/2 (#4695), p-STAT3 (Y705) (#9145), p-STAT3 (S727) (#9134), STAT3 (#9139), Cyclin D1 (#2978) and β-actin (#4967) were purchased from Cell Signaling Technology. Additional antibodies used were GAPDH (#ab8245, Abcam), CCL5 (#ab189841, Abcam) and α-smooth muscle actin (#ab5694, Abcam). Secondary antibodies used were IRDye 800CW Goat anti-Rabbit IgG secondary antibody (#926–32211, LI-COR), IRDye 680RD Goat anti-Mouse IgG secondary antibody (#926–68070, LI-COR), Goat anti-Rabbit IgG Texas Red secondary antibody (#T-2767, ThermoFisher Scientific), Goat anti-mouse IgG Alexa Fluor 488 secondary antibody (#A-11001, ThermoFisher Scientific) and Goat anti-rabbit IgG (H+L) biotinylated secondary antibody (#BA-1000, Vector Labs).