Immunofluorescence analysis of FOLR2 was performed using 70% confluent log phase Raji cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with FOLR2 Polyclonal Antibody (Product # PA5-45768) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cell membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Western blot analysis was performed on whole cell extracts (30 µg lysate) of Raji (Lane 1), Ramos (Lane 2), MCF7 (Lane 3), HT1080 (Lane 4), MDA-MB-231 (Lane 5) and Jurkat (Lane 6). The blot was probed with Anti-FOLR2 Polyclonal Antibody (Product # PA5-45768, 1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4,000 dilution). A 29 kDa band corresponding to FOLR2 was observed across the cell lines tested.