Anti nitrotyrosine rabbit polyclonal antibody

Immunohistochemical analysis was performed as previously described [27 (link),28 (link)], 4 days after DNBS administration. The sections were incubated overnight with primary antibodies anti-ICAM-1 mouse polyclonal antibody (1:100 in Phosphate-buffered saline (PBS), v/v, Santa Cruz Biotechnology SCB, D.B.A, Milan, Italy), anti-P-selectin mouse polyclonal antibody (1:100 in PBS, v/v, SCB, D.B.A, Milan, Italy), anti-PARP mouse polyclonal antibody (1:100 in PBS, v/v, SCB, D.B.A, Milan, Italy), and anti-nitrotyrosine rabbit polyclonal antibody (1:200 in PBS, v/v, Millipore, D.B.A, Milan, Italy). All sections were washed with PBS and then treated as previously reported [29 (link),30 (link)]. Five stained sections from each mouse were scored in a blinded fashion and observed using a Leica DM6 microscope (Leica Microsystems SpA, Milan, Italy) following a typical procedure [31 (link)]. The histogram profile is related to the positive pixel intensity value obtained [32 (link)].

At the end of the experiments, lung tissues were incubated with anti-ICAM-1 murine polyclonal antibody (1/100 in PBS, v/v, Santa Cruz Biotechnology), anti-P-selectin murine polyclonal antibody (1/100 in PBS, v/v, Santa Cruz Biotechnology), anti-PAR murine polyclonal antibody (1/100 in PBS, v/v, Santa Cruz Biotechnology) and anti-nitrotyrosine rabbit polyclonal antibody (1:200 in PBS, v/v, Millipore), as previously described [72 (link),88 (link),103 (link),104 (link),105 (link),106 (link),107 (link),108 (link)]. Immunohistochemical images were collected using (Leica DM6, Milan, Italy) associated with an Imaging system (LasX Navigator, Milan, Italy). The digital images were opened in ImageJ, followed by deconvolution using the color deconvolution plug-in. When the IHC profiler plug-in is selected, it automatically plots a histogram profile of the deconvoluted DAB image, and a corresponding scoring log is displayed. The histogram profile corresponds to the positive pixel intensity value obtained from the computer program. All immunohistochemical analyses were carried out by two observers blinded to the treatment [72 (link),88 (link),103 (link),104 (link),105 (link),106 (link),107 (link),108 (link),109 (link)].

Immunohistochemical analysis was performed as previously described [36 (link),37 (link),38 (link)]. The sections were incubated overnight with primary antibodies: anti-PARP mouse polyclonal antibody (1:100 in PBS, v/v, Santa Cruz Biotechnology (SCB), and anti-nitrotyrosine rabbit polyclonal antibody (1:200 in PBS, v/v, Millipore). Sections were cleaned with PBS and then treated as indicated previously [36 (link)]. Five stained sections from each mouse were scored in a blinded fashion and observed using a Leica DM6 microscope (Leica Microsystems SpA, Milan, Italy) following a typical procedure [39 (link)]. The histogram profile was related to the positive pixel intensity value obtained [40 (link)].