Anti gapdh 0411 sc 47724

Western blot and immunostaining were performed using standard protocols [26 (link)]. Anti-LOX-1 (LOX19-22) sc-66155, anti-AR (N-20) sc-816, anti-GAPDH (0411) sc-47724, anti-NF-kB p65 C-20) sc-372 and anti-IκB-alfa (C-21) sc-371, and anti-STAT3 (c-20) sc-482 antibodies were obtained from Santa Cruz Biotechnology. Anti-p-p65 s536 7F1, anti-p-IκB-alfa S32/S36 5A5, androgen receptor (AR-V7 Specific), and anti-p-STAT3 (Y705) (D3A7) antibodies were obtained from Cell Signaling. For Western blot, the anti-mouse IgG-Alexa Fluor 680 and anti-rabbit IgG-Alexa Fluor 790 secondary antibodies were obtained from Jackson ImmunoResearch. Western blots were analyzed using LICOR-CLX. An anti-rabbit IgG/FITC secondary antibody from Jackson ImmunoResearch was used for immunofluorescences. Photographs were obtained by confocal microscopy (Olympus IX81, Japan). To determine IL-6 concentration from supernatants, the human ELISA Kit (D6050, R&D) was used according to manufacturer’s instructions.

Proteins were prepared as described previously (Cancino et al., 2008 (link)). Tissue protein samples (30 μg) and cellular protein samples (50 μg) were resolved by SDS–PAGE. The immunoblot was carried out using anti-c-Abl (A5844, Sigma-Aldrich, USA), p-c-Abl (Tyr412) (07–788; Millipore), anti-LC3 (NB100-2220), anti-p62 (ab56416), and anti-GAPDH (0411; sc47724; Santa Cruz Biotechnology) antibodies. The secondary antibodies against rabbit or mouse IgGs conjugated with horseradish peroxidase were obtained from Upstate Biotechnology, Lake Placid, NY, United States.

Anti-Rhodopsin antibody (#ab3424) was from Abcam (Cambridge, UK). Rabbit PLD2 (N-term) IgG was from Abgent (San Diego, CA, USA) (#AP14669a). Anti-GAPDH (0411: sc-47724) and m-IgGκ BP-HRP: sc-516102 were from Santa Cruz Biotechnology, Inc. (Dallas, Texas, USA). Anti-PLD1 (#3832), anti-phospho-p70 S6 kinase (Thr389) (#9205) and anti-p70 S6 kinase (#9202) antibodies were from Cell Signaling (Beverly, MA, USA). Polyclonal horse radish peroxidase (HRP)-conjugated sheep anti-mouse IgG (#NA931V) and polyclonal HRP-conjugated donkey anti-rabbit IgG (#NA934V) were purchased from GE Healthcare (Malborough, MA, USA). Alexa Fluor^®488 goat anti-rabbit (#A11008) was from Life Technologies Corporation (Grand Island, NY, USA).