Regulation of Microtubule Acetylation in Cells

The following antibodies were used: CSPGs (2 μg/ml; CC117, EMD Millipore), cycloheximide (10 μg/ml; C0934, Sigma Aldrich), recombinant rat myelin-associated glycoprotein (MAG; 30 μg/ml; P07722, R&D Systems), Y-27632 ROCK inhibitor (10 μM; 1254, Tocris Bioscience), anti- αTAT1 (1:200; ab58742, Abcam), anti-HDAC6 (1:500; NB100-91805, Novus Biologicals), anti-acetylated α-tubulin (1:1000; D20G3, Cell Signaling Technology), anti-α-tubulin (1:5000; DM1A, Sigma-Aldrich), anti-β-actin (1:5000; AC-74, Sigma-Aldrich), anti-β III tubulin (1:5000; MRB₄₃₅P, BioLegend) and anti-GFP (1:500; Sigma-Aldrich). Lentivirus containing GFP (control) or GFP-tagged wild-type αTAT1 constructs, under the human cytomegalovirus (CMV) promoter, was purchased from Dr. Mingjie Li (Washington University School of Medicine, St. Louis, MO; Li et al., 2010 (link)). HDAC6 activity was determined using the fluorometric HDAC6 Activity Assay kit (BioVision), as per manufacturer’s instructions.

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Wong V.S., Picci C., Swift M., Levinson M., Willis D, & Langley B. (2018). α-Tubulin Acetyltransferase Is a Novel Target Mediating Neurite Growth Inhibitory Effects of Chondroitin Sulfate Proteoglycans and Myelin-Associated Glycoprotein. eNeuro, 5(1), ENEURO.0240-17.2018.

Publication 2018

CC117 ab58742 AC-74 MRB435P anti-GFP fluorometric HDAC6 Activity Assay kit

Actin Antibodies Assay Biologicals Cycloheximide Fluorometric Human cytomegalovirus Lentivirus Myelin associated glycoprotein Novus Tubulin Y 27632 α tubulin

Corresponding Organization :

Other organizations : Burke Medical Research Institute, University of Waikato, Cornell University

Top 5 similar protocols

Variable analysis

independent variables

CSPGs (2 μg/ml)
Cycloheximide (10 μg/ml)
Recombinant rat myelin-associated glycoprotein (MAG; 30 μg/ml)
Y-27632 ROCK inhibitor (10 μM)
Lentivirus containing GFP (control) or GFP-tagged wild-type αTAT1 constructs, under the human cytomegalovirus (CMV) promoter

dependent variables

HDAC6 activity

control variables

Anti- αTAT1 (1:200)
Anti-HDAC6 (1:500)
Anti-acetylated α-tubulin (1:1000)
Anti-α-tubulin (1:5000)
Anti-β-actin (1:5000)
Anti-β III tubulin (1:5000)
Anti-GFP (1:500)

positive controls

Lentivirus containing GFP (control)

negative controls

Not explicitly mentioned

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