Goat anti lyve1 af2125

Deparaffinization and rehydration of paraffin sections (1 µm) was performed according to standard protocols. Antigen retrieval was carried out with epitope retrieval solution (Zytomed Systems, Germany) at pH 6. The first antibody was incubated over night at 4 °C, the secondary antibody was applied for 1 h at room temperature after three washing steps with PBS. Sections were mounted with Dako fluorescent mounting medium (Dako, Agilent technologies, USA) and photographed with ECLIPSE Ni-E microscope (Nikon). Antibodies: goat anti-Lyve1 (AF2125, R&D Systems, USA), rat anti-Endomucin (14-5851-82, Thermo Fisher Scientific, USA), rabbit anti-glutamine synthetase (G2781, Sigma-Aldrich, Germany), goat anti-Arginase I (sc-18351, Santa Cruz, USA), goat anti-RhBg (PA5-19369, Thermo Fisher Scientific, USA), Alexa-Fluor 488, Alexa-Fluor 647 and Cy3-conjugated secondary antibodies were purchased from Dianova (Germany).

Primary antibodies: Rabbit anti-cleaved Caspase 3 (9661S, Cell Signaling Technology, Danvers, USA), rabbit anti-Ki-67 (ab16667, Abcam, Cambridge, UK), rat anti-CD31 (DM3614P, Dianova, Hamburg, Germany), goat anti-CD32b (AF1460, R&D Systems, Minneapolis, USA), goat anti-Lyve1 (AF2125, R&D Systems, Minneapolis, USA), rat anti-Endomucin (14–5851–82, Thermo Fisher Scientific, Waltham, USA), rabbit anti-TRP-2 (ab74073, Abcam, Cambridge, UK), rabbit anti-wide spectrum cytokeratin (ab9377, Abcam, Cambridge, UK), goat anti-alpha smooth muscle actin (ab21027, abcam, Cambridge, UK), rat anti-CD45 (550539, BD Biosciences, Franklin Lakes, USA), rat anti-CD68 (137002, BioLegend, San Diego, USA), rabbit anti-Melan A (NBP1-30151, Novus, Minneapolis, USA). For Western blotting: rabbit anti-Akt (9272S, Cell Signaling Technology, Danvers, USA), rabbit anti-phospho-Akt (9271S, Cell Signaling Technology, Danvers, USA). Secondary antibodies: Alexa-Fluor 488, Alexa-Fluor 647 and Cy3-conjugated secondary antibodies were purchased from Dianova (Hamburg, Germany). For Western Blotting a rabbit anti-IgG HRP conjugated antibody was used. (Merck, Darmstadt, Germany).