Quantifying Membrane Protein Expression

Whole cell extracts were prepared in 1% NP-40 lysis buffer containing protease and phosphatase inhibitors as described (8 (link)). Membranes were immunolabeled with anti-GLUT4 (sc-1608, Santa Cruz Biotech), anti-myc (9E10) (sc-40, Santa Cruz Biotech), anti-CRISPLD2 (NBP1–85143, Novus Biologicals) anti-adiponectin (Philipp Scherer) or anti-tubulin antibody (2148, Cell Signaling) and visualized with secondary antibodies conjugated with AlexaFluor 680 (Invitrogen). Fluorescence was quantified with a Li-Cor Odyssey imager (Li-Cor Biosciences, USA).

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Jackson R.M., Griesel B.A., Short K.R., Sparling D., Freeman W.M, & Olson A.L. (2019). Weight loss resulted in increased expression of the anti-inflammatory protein, CRISPLD2, in mouse adipose tissue. Obesity (Silver Spring, Md.), 27(12), 2025-2036.

Publication 2019

anti-myc (9E10) (sc-40, AlexaFluor 680 Li-Cor Odyssey imager

Adiponectin Anti antibody Antibodies Biologicals Buffer Cell extracts Fluorescence Glut4 Inhibitors Membranes Novus Np 40 Phosphatase Protease Tubulin

Corresponding Organization : University of Oklahoma Health Sciences Center

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Variable analysis

independent variables

Whole cell extracts were prepared in 1% NP-40 lysis buffer containing protease and phosphatase inhibitors

dependent variables

Fluorescence was quantified with a Li-Cor Odyssey imager (Li-Cor Biosciences, USA)

control variables

Anti-tubulin antibody (2148, Cell Signaling)

controls

Positive control: anti-tubulin antibody (2148, Cell Signaling)
Negative controls: not explicitly mentioned

Annotations

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