Western blot analysis of protein translocation

Panel A. A549 cells (2 x 10^6) were either mock-treated (-) or incubated (+) with 20µg/mL TNFα for 20 minutes and fractionated using the Subcellular Protein Fractionation Kit. Each extract (10µg) was analyzed by Western blot using an anti-NFκB p65 antibody. Panel B. Serum-starved HeLa cells (2 x 10^6) were either mock-treated (-) or incubated (+) with 1µM PMA for 20 minutes and fractionated. Each extract (10µg) was analyzed by Western blot using an anti-PKCα antibody. Goat anti-rabbit or anti-mouse (H+L) HRP was used as the secondary antibody, and SuperSignal West Dura Chemiluminescent Substrate (Part No. 34076) was used for detection.

Panel A. A549 cells (2 x 10^6) were either mock-treated (-) or incubated (+) with 20µg/mL TNFα for 20 minutes and fractionated using the Subcellular Protein Fractionation Kit. Each extract (10µg) was analyzed by Western blot using an anti-NFκB p65 antibody. Panel B. Serum-starved HeLa cells (2 x 10^6) were either mock-treated (-) or incubated (+) with 1µM PMA for 20 minutes and fractionated. Each extract (10µg) was analyzed by Western blot using an anti-PKCα antibody. Goat anti-rabbit or anti-mouse (H+L) HRP was used as the secondary antibody, and SuperSignal West Dura Chemiluminescent Substrate (Part No. 34076) was used for detection.

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