Ab134185

In western blotting, cells were lysed in 1× SDS loading buffer (50 mM Tris-HCl pH6.8, 10% glycerol, 2% SDS, 0.05% bromophenol blue, and 1% 2-mercaptoethanol). Antibodies were listed as follows: anti-ZBTB38 antibody (21906-1-AP, Proteintech), anti-DKK1 (21112-1-AP, Proteintech), anti-PRKDC (SC-5282, Santa Cruz), anti-HA (51064-2-AP, Proteintech), anti-FLAG (20543-1-AP, Proteintech), anti-GAPDH (10494-1-AP, Proteintech), and anti-TUBULIN (ab134185, Abcam).
We did immunoblot as previously described [22 (link)]. In brief, all proteins were separated by SDS–PAGE and were transferred to polyvinylidene difluoride membranes (Millipore). Horseradish peroxidase-labeled secondary antibodies and an enhanced chemiluminescence system were used for signal detection. Protein was visualized using ChemiDoc XRS chemiluminescence detection and imaging system (Bio-Rad Laboratories) or KODAK film machine.

Cells were lysed using 1× SDS loading buffer (50 mM Tris-HCl pH 6.8, 10% glycerol, 2% SDS, 0.05% bromophenol blue, and 1% 2-mercaptoethanol). Antibodies were listed as follows: anti-HNF1B antibody (12533-1-AP, Proteintech), anti-E-cadherin (610181, BD Transduction Laboratories), anti-SLUG (C19G7, Cell Signaling Technology), anti-SNAI1 (C15D3, Cell Signaling Technology), anti-EZH2 (AC22, Cell Signaling Technology), anti-HA (51064-2-AP, Proteintech), anti-FLAG (20543-1-AP, Proteintech), anti-RBBP7 (20365-1-AP, Proteintech), and anti-tubulin (ab134185, Abcam). For immunoblot, proteins were separated by SDS–PAGE and transferred to polyvinylidene difluoride membranes (Millipore). HRP-conjugated secondary antibodies (Jackson laboratories) and enhanced chemiluminescence system was used for signal detection. Protein was visualized using KODAK film machine or ChemiDoc XRS chemiluminescence detection and imaging system (Bio-Rad Laboratories).