Fig 1: Platelet neutrophil complex multiplex immunohistochemistry in pig lung and liver tissues. To ensure colour detection systems were functional, separate sections were immunostained with either anti-CD42b shown in red (A) or anti-MPO shown in blue. Representative images of neutrophil immunohistochemistry (anti-CD42b) in pig lung tissue (C,D) and pig liver tissue (E,F) are also shown. Asterisks denote blood vessels. Arrows point to PNCs. Scale bar = 50 µm in Figures (A,B) (×40 objective), 100 µm in Figure (C,E) (×20 objective) and 30 µm in Figures (C,E) (×63 objective) for neutrophil complex multiplex immunohistochemistry in pig lung and liver tissues. Formalin fixed paraffin wax embedded (FFPE), and pig tissues were prepared and immunostained with anti-myeloperoxidase (MPO) antibody (ab9535, Abcam, Cambridge, UK) and anti-CD42b antibody (ab183345, Abcam, Cambridge, UK), prepared at 1/50 and 1/100 dilutions, respectively. Platelets appear red, and neutrophils appear blue. To ensure colour detection systems were functional, separate sections were immunostained with either anti-CD42b shown in red (A) or anti-MPO shown in blue. Representative images of neutrophil immunohistochemistry (anti-CD42b) in pig lung tissue (C,D) and pig liver tissue (E,F) are also shown. Asterisks denote blood vessels. Arrows point to PNCs. Scale bar = 50 µm in Figures (A,B) (×40 objective), 100 µm in Figure (C,E) (×20 objective) and 30 µm in Figures (C,E) (×63 objective).
Fig 2: Platelet neutrophil complex multiplex immunohistochemistry in rat lung and liver tissues. Frozen rat tissues were prepared and immunostained with anti-myeloperoxidase (MPO) antibody (ab9535, Abcam, UK) and anti-CD61 antibody (PB9647, Boster, USA), prepared at 1/25 and 1/100 dilutions, respectively. Platelets appear red, and neutrophils appear blue. Representative images of platelet neutrophil complex (PNC) staining in rat lung tissue (A,B) and rat liver tissue (C,D). Arrows denote CD61+ MPO+ events. Scale bar = 100 µm in Figures (A,B) (×20 objective) and 30 µm in Figures (B,C) (×63 objective).
Fig 3: Confocal imaging of SH activity in relation to myeloperoxidase (MPO), a marker for neutrophils. Sections went through the tissue-ABPP protocol to label SHs (red) and were thereafter immunostained for MPO (yellow), followed by DAPI staining to visualize nuclei (blue). Panel a shows overall staining pattern throughout the coronal section plane. A control section undergoing identical staining protocol with no primary antibody is illustrated at top. Panel b shows staining pattern in glioma region characterized by intense SH activity originating from individual cells (TAMRA-FP hotspots). Panel c shows staining pattern in glioma region characterized by intense SH activity originating from cell clusters (TAMRA-FP hotspot clusters). Panel d shows staining pattern in healthy brain (cortex). Note detectable expression of MPO in the glioma (a). Note MPO-positive cells in the region of TAMRA-FP hotspots and marked co-localization of MPO with the TAMRA-FP signal (b). Note abundance of MPO-positive cells within TAMRA-FP hotspot clusters, as well as close match of MPO staining with TAMRA-FP hotspot clusters (c). MPO is not visible in control cortical region (d). Primary antibody rabbit anti-MPO (Abcam, cat# ab9535), dilution 1:25, secondary antibody Goat anti-rabbit IgG-Alexa Fluor 647 conjugate, dilution 1:100. Sections were from female rat 11. Scale bars: 1 mm in a, 20 µm in b-d. Images were adjusted for brightness and contrast
Supplier Page from Abcam for Anti-Myeloperoxidase antibody