mouse serum dilutions  (Jackson Immuno)

Journal: PLoS ONE

Article Title: Platelet-Activating Factor Induces TLR4 Expression in Intestinal Epithelial Cells: Implication for the Pathogenesis of Necrotizing Enterocolitis

doi: 10.1371/journal.pone.0015044

Figure Lengend Snippet: IEC-6 cells were treated without (a,b,c), or with (d,e,f) 2 µM PAF for 1 h then fixed and processed for STAT3 immunohistochemistry (a,d; c and f red) and nuclei were counterstained with Hoechst dye (b,e; c and f blue). COS-7 cells were first incubated with AD-PAFR for 24 hrs then without (g,h), or with (i,j) 1 µM PAF for 1 h. Triple label immunohistochemistry was performed with anti STAT3 (red), anti PAFR (green) and the DNA binding dye Hoechst (blue). Rat small intestinal cells (IEC-6) were treated as indicated, then Western blots were performed on cytoplasmic and nuclear fractions using antibodies against STAT3 and phorphorylated STAT3 (p-STAT3).

Article Snippet: Following washing and blocking, cells were probed with mouse monoclonal anti-STAT3 (diluted 1∶50 in 10% goat serum) (Invitrogen/Molecular Probes, Carlsbad, CA) overnight at 4°C in a humidified chamber followed by incubation with Alexa Fluor 546 labeled goat anti-mouse secondary antibody for 1 h at 37°C in a humidified chamber.

Techniques: Immunohistochemistry, Incubation, Binding Assay, Western Blot