LDB1 Western Blot
Description
Western blot validating auxin-inducible degron system for LDB1. Western blot for LDB1 (GAPDH as loading control) in LDB1-AID G1E-ER4 cells. 3 samples (parental G1E-ER4 cells, LDB1-AID clone 1, LDB1-AID clone 2); 2 treatment conditions (untreated, 4hr auxin treated). Hypothesis - auxin treatment will deplete LDB1 protein levels in LDB1-AID clones, but not parental cells. Sample order from left to right: parental untreated, parental 4hr auxin, LDB1-AID clone 1 untreated, LDB1-AID clone 1 4hr auxin, LDB1-AID clone 2 untreated, LDB1-AID clone 2 4hr auxin.
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Steps to reproduce
Western blot analysis was performed for LDB1 (Thermo Fisher Scientific, PA5-56948) in both LDB1-AID clonal lines and parental G1E-ER4 cells in the absence of auxin and in 4 hour auxin treatment conditions. Samples were lysed in complete RIPA lysis buffer and sonicated with the Bioruptor Pico (Diagenode, 3 min: 30sec on, 30sec off, ‘easy’ mode. Protein lysates were run on a 4-12% Bis-Tris gel. GAPDH was used as a loading control (Santa Cruz Biotechnology, sc-32233).