For detection of ubiquitinated and SUMOylated TOP3Accs,we performed DUST assay as described previously22 (link). Briefly, nucleic acids and covalent protein-nucleic acid adducts were recovered from FLAG-tagged R364W-TOP3A transfected cells using the RADAR assay. 10 μg of each RADAR assay sample was digested with 250 units benzonase nuclease (EMD Millipore, 100 units/μl) in the presence of 5 mM CaCl2, followed by SDS-PAGE electrophoresis for immunodetection of total TOP3Accs and ubiquitinated and SUMOylated TOP3Accs by probing with rabbit anti-TOP3A (dilution 1:1000, Proteintech, Rosemont, IL, CAT#: 14525-1-AP), mouse anti-ubiquitin antibody (dilution 1:500, Cell Signaling Technology, Cat# 3936), rabbit anti-SUMO-1 (dilution 1:1000, Cell Signaling Technology, Cat# 4940) and rabbit anti-SUMO-2/3 antibody (dilution 1:1000, Cell Signaling Technology, Cat# 4971), respectively.
Mouse anti ubiquitin antibody
Manufactured by Cell Signaling Technology
Mouse anti-ubiquitin antibody is a primary antibody that specifically recognizes ubiquitin, a small regulatory protein found in eukaryotic cells. This antibody can be used to detect and study the expression and distribution of ubiquitin in various biological samples.
Automatically generated - may contain errors
2 protocols using mouse anti ubiquitin antibody
1
Detection of Ubiquitinated and SUMOylated TOP3A
2
Antibody Validation for Hepatitis B Protein Detection
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The rabbit anti-20S (PSMB9) polyclonal antibody (14544-1-AP) and rabbit anti-26S (PSMD2) polyclonal antibody (14748-1-AP) were purchased from Proteintech (Chicago, United States). The rabbit anti-mTOR (7C10) monoclonal antibody (#2983T), anti-Flag antibody (#14793S), and mouse anti-Ubiquitin antibody (#3936) were acquired from Cell Signaling Technology (CST, Boston, United States). The rabbit anti-HBcAg (B0586) was purchased from Dako (Copenhagen, Denmark). The Rabbit IgG (NI01) was acquired from Millipore (Darmstadt, Germany) and the mouse anti-HBxAg (1844) was purchased from ViroStat (ME, United States). The mouse anti-GAPDH (1A6) monoclonal antibody (MB001) was obtained from Bioworld (Bloomington, United States). The rabbit anti-HBsAg (NB100-62652) was purchased from Novus (CO, United States). The IPKine™ HRP anti-mouse IgG LCS antibody was obtained from Abbkine (Wuhan, China).
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