Western Blot Analysis of UCP2 Expression

Western blots were performed with protein samples as indicated. After transfer to a nitrocellulose membrane samples were incubated with either an antibody directed against UCP2 and subsequently with an antibody directed against actin or voltage-dependent anion exchanger (VDAC) or GAPDH. Signals were detected by a fluorescence coupled secondary antibody (Abcam, ab6940). Western blots dealing with UCP2 expression were performed with an antibody evaluated before.¹⁸ (link) Specificity was tested in tissue samples from spleens of UCP2⁻^/⁻ mice and wild-type mice, respectively (Supplementary material online, Figure S1). Control peptide for UCP2 was a recombinant human UCP2.¹⁹ (link)

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Esfandiary A., Kutsche H.S., Schreckenberg R., Weber M., Pak O., Kojonazarov B., Sydykov A., Hirschhäuser C., Wolf A., Haag D., Hecker M., Fink L., Seeger W., Ghofrani H.A., Schermuly R.T., Weißmann N., Schulz R., Rohrbach S., Li L., Sommer N, & Schlüter K.D. (2019). Protection against pressure overload-induced right heart failure by uncoupling protein 2 silencing. Cardiovascular Research, 115(7), 1217-1227.

Publication 2019

ab6940

Actin Anion Antibody Fluorescence Gapdh Human Membrane Mice Nitrocellulose Peptide Protein Tissue Western blots

Corresponding Organization : University of Giessen

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Variable analysis

independent variables

Presence or absence of UCP2 protein (UCP2-/- mice vs. wild-type mice)

dependent variables

UCP2 protein expression
Actin protein expression
VDAC protein expression
GAPDH protein expression

control variables

Western blot protocol (sample preparation, transfer, incubation with primary and secondary antibodies, detection method)

controls

Positive control: Recombinant human UCP2 protein
Negative control: Tissue samples from UCP2-/- mice

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