Quantifying APP and Ubiquilin-1 Interactions

L-685,458 and puromycin were purchased from Sigma. β-Actin antibody (1:10,000) was purchased from Sigma. The APP C-terminal antibody (A8717; 1:1,000) was purchased from Sigma. The ubiquilin-1 antibody (1:160) was purchased from Zymed. The HRP-conjugated secondary antibodies (anti-mouse and anti-rabbit) (1:10,000) were purchased from GE. The plasmids APP-Gal4, Gal4-UAS-luciferase (encoding firefly luciferase) were kindly provided from Dr. Thomas Südof, and have been described (Cao and Sudhof, 2001 (link)). The previously reported ubiquilin-1 expressing plasmid was constructed from the pCMV vector and was kindly provided by Dr. Mervyn J. Monteiro (Mah et al., 2000 (link)).

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Zhang C., Inamdar S.M., Swaminathan S., Marenda D.R, & Saunders A.J. (2022). Association of the Protein-Quality-Control Protein Ubiquilin-1 With Alzheimer’s Disease Both in vitro and in vivo. Frontiers in Neuroscience, 16, 821059.

Publication 2022

L-685,458 puromycin β-Actin antibody APP C-terminal antibody (A8717 HRP-conjugated secondary antibodies (anti-mouse and anti-rabbit)

Actin Antibodies Antibody Firefly luciferase L 685 458 Luciferase Mouse Plasmid Puromycin Rabbit Vector

Corresponding Organization : Drexel University

Other organizations : National Science Foundation, Division of Biological Infrastructure

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Variable analysis

independent variables

L-685,458
Puromycin
APP-Gal4
Gal4-UAS-luciferase
Ubiquilin-1 expressing plasmid

dependent variables

Firefly luciferase activity

control variables

β-Actin antibody (1:10,000)
APP C-terminal antibody (A8717; 1:1,000)
Ubiquilin-1 antibody (1:160)
HRP-conjugated secondary antibodies (anti-mouse and anti-rabbit) (1:10,000)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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