Polyclonal goat anti rabbit secondary antibody sc 66931

Western blot analysis was performed as described previously [61 (link)], using the following antibodies against rabbit polyclonal CD36 (Origene, TA500921S; 1:2000): rabbit polyclonal ABCA1 (Santa Cruz Biotecnology, Sc-58219; 1:300), rabbit polyclonal LXRα (Origene, TA805015S; 1:2000), rabbit polyclonal PPARγ (Millipore, ABN1445; 1:500) and mouse monoclonal β-actin (Santa Cruz Biotechnology, Inc., Dallas, TX, USA). The blots were then incubated for 1 h at room temperature with goat anti-mouse secondary antibody (Sc-2005; 1:2000; Santa Cruz Biotechnology) or polyclonal goat anti-rabbit secondary antibody (Sc-66931; 1:5000; Santa Cruz Biotechnology). The nitrocellulose was scanned using a computerized densitometric system (Bio-Rad Gel Doc 1000, Milan, Italy). Protein levels were normalized to the housekeeping proteins β-actin to adjust for variability in protein loading and expressed as a percentage of vehicle control.

Western blot analysis was performed as described previously [16 (link)] using the following antibodies against rabbit polyclonal Bcl-2 (Sc-492; 1:500), rabbit polyclonal cleaved caspase-3 (Sc-22171-R; 1:400), rabbit polyclonal Bax (Sc-493; 1:500), rabbit polyclonal pAkT (sc–135650; 1:1000), rabbit polyclonal p-PI3-kinase (sc-12929; 1:500) and mouse monoclonal β-actin (Santa Cruz Biotechnology, Inc., Dallas, TX, USA). The blots were then incubated for 1 h at room temperature with goat anti-mouse secondary antibody (Sc-2005; 1:2000; Santa Cruz Biotechnology) or polyclonal goat anti-rabbit secondary antibody (Sc-66931; 1:5000; Santa Cruz Biotechnology).
To verify if the U937-PMA driven differentiation was also associated with the expression of macrophage-selective markers, we performed Western blot analyses with the following monoclonal antibodies: anti-CD206 (monoclonal, 1:1000, Santa Cruz), anti-CD14 (monoclonal, Santa Cruz). The differentiation of SH-SY5Y cells was detected using the following antibodies: anti-Nestin (monoclonal, 1:1000, Santa Cruz), rabbit anti-MAP2 (policlonal, 1:1000, Millipore) (Figure S1).