Immunofluorescence analysis of KPNA2 was performed using HEK cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with KPNA2 Recombinant Rabbit Monoclonal Antibody (Product # MA5-32761) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11006) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing cytoplasmic localization of KPNA2. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Western blot was performed using Anti-Importin subunit alpha-1 (KPNA2) Monoclonal Antibody (Product # MA5-32761) on whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), A549 (Lane 2), HeLa (Lane 3), NIH/3T3 (Lane 4), PC-12 (Lane 5), LNCaP (Lane 6), Mouse Testis (Lane 7), Mouse Thymus (Lane 8), Mouse colon (Lane 9), Mouse Spleen (Lane 10) and Rat Testis (Lane 11). A 57.86 kDa band corresponding to Importin subunit alpha-1 (KPNA2) was observed across samples tested except Mouse colon which is reported to be negative. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1,000 dilution) and detected by Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).