Alexa fluor 488 goat anti guinea pig igg h l

Immunostaining was performed as previously described^{17 (link)}. Primary antibodies were: guinea pig anti-VGLUT1 (135304, SYSY), rabbit anti-VGAT (131013, SYSY), and neuronal class III beta-tubulin antibody (clone TUJ1) (Covance, MMS-435P, Princeton, NJ, USA) diluted 1:250. Secondary antibodies were: Alexa Fluor 488 goat anti-guinea pig IgG (H + L) (A11073, Life Technologies, Carlsbad, CA, USA), and Alexa Fluor 568 goat anti-rabbit IgG (H + L) (A11036, Life Technologies, Carlsbad, CA, USA). All antibodies were diluted 1:500 except where otherwise specified. Nuclei were stained with Hoechst dye (3 μg/mL) (B2261, Sigma Aldrich, Milan, Italy). Images were acquired with a Leica SP8 confocal microscope and analyzed with ImageJ.

The following primary antibodies were used in this study: CD26 mouse immunoglobulin (IgG) 2a (BD Pharmingen, San Jose, CA, United States); CD31 mouse IgG1 (BD Pharmingen); cytokeratin 19 mouse IgG2b (CK19; Progen, Heidelberg, Germany); GST-p rabbit IgG (MBL CO., LTD, Nagoya, Japan); HNF4a goat IgG (Santa Cruz Biotechnology, Dallas, TX, United States); hepatic sinusoidal endothelial cell mouse IgG2a (SE-1; IBL CO., LTD, Gunma, Japan); human vimentin mouse immunoglobulin (IgG) (DAKO: Agilent technologies, Santa Clara, CA, United States); human albumin (SIGMA) mouse IgG2a; human CD31 (DAKO) mouse IgG1; human CK19 (DAKO) mouse IgG1; and CK19 mouse IgG2b (Progen, Heidelberg, Germany). The second antibodies used were Alexa Fluor 488 goat anti-mouse IgG1(γ1) (Invitrogen: Thermo Fisher Scientific, Waltham, MA, United States), Alexa Fluor 647 goat anti-mouse IgG1 (γ1) (Invitrogen), Alexa Fluor 555 goat anti-mouse IgG2a (γ2a) (Invitrogen), Alexa Fluor 488 goat anti-mouse IgG2b (γ2b) (Invitrogen), Alexa Fluor 647 goat anti-rabbit IgG (H+L) (Invitrogen), and Alexa Fluor 488 goat anti-guinea pig IgG (H+L) (Life technologies: Thermo Fisher Scientific, Waltham, MA, United States).