Biochemical Characterization of Ion Channels

C20:4-Gly-d₈, C20:4-Ser, GSK1016790A, PGE₂, and the COX (ovine) Colorimetric Inhibitor Screening Assay Kit were purchased from Cayman Chemical Company; capsaicin, glutamine, phenylalanine, and oleic acid were purchased from Sigma; 2-APB was purchased from Santa Cruz Biotechnology; and C20:4-Phe was purchased from Abcam. The synthesis of C18:1-Phe, C18:1-Leu, C18:1-Gln, and C18:1-Lys has been described previously (8 (link)). Plasmids were obtained from the following sources: C-terminally tagged mouse TRPV1 (catalog no. MR227160; OriGene), C-terminally tagged human TRPV3 (catalog no. RC211184; OriGene); C-terminally tagged rat TRPV4 (plasmid no. 45751; Addgene); and pTurboRFP-Mito (catalog no. FP237; Evrogen). Mouse TRPA1 was a generous gift from Jorg Grandl, Duke University, Durham, NC. C2C12 cells and HEK293A cells were obtained from ATCC and Thermo Fisher Scientific, respectively.

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Long J.Z., Roche A.M., Berdan C.A., Louie S.M., Roberts A.J., Svensson K.J., Dou F.Y., Bateman L.A., Mina A.I., Deng Z., Jedrychowski M.P., Lin H., Kamenecka T.M., Asara J.M., Griffin P.R., Banks A.S., Nomura D.K, & Spiegelman B.M. (2018). Ablation of PM20D1 reveals N-acyl amino acid control of metabolism and nociception. Proceedings of the National Academy of Sciences of the United States of America, 115(29), E6937-E6945.

Publication 2018

GSK1016790A COX (ovine) Colorimetric Inhibitor Screening Assay Kit capsaicin glutamine phenylalanine oleic acid 2-APB C2C12 cells HEK293A

Assay Capsaicin Cayman Cells Colorimetric Gsk1016790a Human Mito Mouse Oleic acid Ovine Pge2 Plasmid Synthesis Trpv4

Corresponding Organization : Harvard University

Other organizations : University of California, Berkeley, Scripps Research Institute, Brigham and Women's Hospital, Beth Israel Deaconess Medical Center

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Variable analysis

independent variables

C20:4-Gly-d₈
C20:4-Ser
GSK1016790A
PGE₂
Capsaicin
Glutamine
Phenylalanine
Oleic acid
2-APB
C20:4-Phe
C18:1-Phe
C18:1-Leu
C18:1-Gln
C18:1-Lys

dependent variables

Outcomes measured using the COX (ovine) Colorimetric Inhibitor Screening Assay Kit

control variables

C-terminally tagged mouse TRPV1
C-terminally tagged human TRPV3
C-terminally tagged rat TRPV4
PTurboRFP-Mito
Mouse TRPA1
C2C12 cells
HEK293A cells

Annotations

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