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HRI Antibody (702551) in WB
Western blot analysis was performed on Membrane extracts (30 µg lysate) of K-562 (Lane 1), K-562 treated with H2O2 (150 uM for 1 hr)(Lane 2), HEL 92.1.7 (Lane 3), HEL 92.1.7 treated with H2O2 (150 uM for 1 hr) (Lane 4), T-47D (Lane 5), T-47D treated with H2O2 (150 uM for 1 hr) (Lane 6), HEK-293 (Lane 7) and HEK-293 treated with H2O2 (150 uM for 1 hr) (Lane 8). The blots were probed with ABfinity? Anti-eIF2AK1 / Heme regulated inhibitor (HRI) Rabbit Monoclonal Antibody (Product ## 702551, 1-2 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal? Secondary Antibody, HRP conjugate (Product ## A27036, 0.4 µg/ml, 1:2500 dilution). A 71 kDa band corresponding to eIF2AK1 / Heme regulated inhibitor (HRI) was observed across the cell lines (with treatment difference) tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product ## NP0321BOX), XCell SureLock? Electrophoresis System (Product ## EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product ## LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product ## IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product ## WP20005).
