Goat anti mouse alexa fluor 790

On-cell western [74 ] was performed with U2OS MYC-ER cells. 72,000 cells were plated in each well of a 24-well plate, and 24 hours later, treated ± 4OHT for 48 hours. Cells were then fixed with 3.7% paraformaldehyde (Sigma) but not permeabilized. Wells were washed with tris-buffered saline, and stained with primary antibodies mouse anti-LAT1 BU53 (Novus NBP2-50465AF647) or mouse IgG2A isotype control (Novus IC003R), and secondary antibodies goat anti-mouse Alexa Fluor 790 (Thermo Scientific A11357) or CellTag 700 Stain (Licor 926–41090), which is used to quantify total cell number and intensity. The stained plate was then digitally imaged using a Licor Odyssey CLx infrared imager (Licor), and well intensities in both channels (700 nM for CellTag, 800 for LAT1 or isotype control) were quantified using ImageStudio software (Licor). Individual wells were treated as biological replicates, and results are representative of two separate experiments.

The primary antibodies used in this study are as following: anti-AtCRY1 (1:5,000, homemade) (Lin et al., 1996 (link)), anti-AtCRY2 (1:3,000, homemade) (Lin et al., 1998 (link)), anti-HSP90 (1:20,000, cat # AbM51099-31-PU, Beijing Protein Innovation), anti-GFP (1:2,000, cat # 598, MBL), anti-Myc (1:2,000, cat # 05-724, Millipore), anti-Flag (1:1,500, cat # F3165, Sigma-Aldrich).
The secondary antibodies used in this study are as following: anti-Mouse-HRP (1:15,000, cat # 31430, Thermo Fisher Scientific), anti-Rabbit-HRP (1:15,000, cat # 31460, Thermo Fisher Scientific), goat anti-mouse Alexa Fluor 790 (1:15,000, cat # A11357), goat anti-rabbit IgG-Alexa Fluor 790 (1:15,000, cat # A11369), goat anti-mouse IgG-Alexa Fluor 680 (1:15,000, cat # A21058), goat anti-rabbit IgG-Alexa Fluor 680 (1:15,000, cat # A21109).