Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor® Plus 800 conjugate for Western Blot
Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A32735) in WB
Western blot analysis of Heat Shock Protein 90 (Hsp90) and histone deacetylase 1 (HDAC1) was performed by loading 3-fold serial dilutions of A431 whole cell lysate (starting at 15ug) and 2ul of the PageRuler Prestained NIR Protein Ladder (Product # 26635) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to Nitrocellulose Membranes (Product # 88018) and blocked with Fluorescence Blocker for 30 min. Membranes were probed with a Hsp90 polyclonal antibody (Product # PA3-013) at a dilution of 1:5000 and an HDAC1 polyclonal antibody at a dilution of 1:5000 (Product # PA1-1110) overnight at 4°C on a rocking platform, washed with TBST, and probed with an Invitrogen Alexa Fluor Plus 800 Goat anti-Rabbit IgG secondary antibody (Product # A32735) at dilutions of 1:40,000 for 45 minutes. Blots were imaged on an Infrared fluorescence imaging system.