Goat anti mouse dylight 633

Antibodies used for flow cytometry: Alexa Fluor 488 Mouse anti-STAT3 (pY705) (Clone 4/P-STAT3), BD Phosflow, Franklin Lakes, NJ, USA, Catalog No. 557814, Lot: 4003621; PE Mouse anti-STAT3 (Clone M59-50), BD Phosflow, Catalog No. 560391, Lot: 7046704; Mouse anti-gp130 (Clone BR-3), Hölzel Diagnostics, Cologne, Germany, Catalog No. 852.060.000, Lot:P11016D6.
Antibodies used for immunodetection of Western Blots: Rabbit anti-STAT3 (pY705) (Clone D3A7), Cell Signaling Technology, Cambridge, UK, Catalog No. 9145, Lot: 34; Rabbit anti-STAT3 (pS727) (Polyclonal), Cell Signaling Technology, Catalog No. 9134, Lot: 21; Mouse anti-STAT3 (Clone 124H6), Cell Signaling Technology, Catalog No. 9139, Lot: 10; Rabbit anti-SOCS3 (Clone C204), Immuno-Biological Laboratories, Fujioka, Japan, Catalog No. 18391, Lot: 0G-901; Mouse anti-HSC70 (Clone 1F2-H5), StressMarq Biocsciences, Victoria, Canada, Catalog No. SMC-151, Lot: 0706; Goat anti-Mouse DyLight 633 (polyclonal), Thermo Fisher Scientific, Catalog No. 815-968-0747, Lot: NB167398; Goat anti-Rabbit DyLight 550 (polyclonal), Thermo Fisher Scientific, Catalog No. 815-968-0747, Lot: NB165012.

The OCT 12‐μm‐thick frozen sections were prepared on glass slides (Ultra Superfrost Plus, Thermo Fisher Scientific, Waltham, Massachusetts, USA) and further subjected to an immunofluorescence protocol, with minor modifications.²⁹ After post‐fixation and incubation in the blocking solution (BS), sections were incubated with a combination of primary antibodies: rabbit polyclonal anti‐CBR1, rabbit polyclonal anti‐CBR2, mouse monoclonal anti‐pan‐cytokeratin, mouse monoclonal anti‐vimentin, and chicken polyclonal anti‐PGP 9.5 overnight at 4°C. The next day, sections were incubated with goat‐anti‐rabbit DyLight488, goat‐anti‐chicken DyLight594, and goat‐anti‐mouse DyLight633 (all primary and secondary antibodies purchased from Thermo Fisher Scientific, Waltham, Massachusetts, USA) for 2 hours at room temperature (RT) as well as counterstained with DAPI (Santa Cruz Biotechnology, Santa Cruz, California, USA). The protocol described above also was used for in vitro analysis. Fluorescence intensity (FI) analysis was performed as previously described with minor modification using Fiji‐ImageJ Software (Fiji‐ImageJ, National Institute of Health, Bethesda, Maryland, USA).²⁹, ³⁰