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2 protocols using carnitine palmitoyltransferase 1a

1

Striatal Protein Expression Analysis

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Striatum samples were homogenized by sonication in SDS buffer (10%). After determining the dynamic range of detection for each antibody, 20 μg of total protein were resolved in 4-20% SDS-PAGE and then transferred onto polyvinylidene difluoride (PVDF) membranes using the Trans-Blot Turbo transfer system (Bio-rad). Briefly, membranes were incubated overnight at 4°C with primary antibodies of interest anti-: TH (Cell Signaling; s58844), mTOR (Cell Signaling; s2983), p-mTOR (Cell Signaling; s5536), iNOS (Cell Signaling; s13120), MPC1 (Cell Signaling; s14462), pyruvate dehydrogenase (Cell Signaling; s3205), p-pyruvate dehydrogenase (Cell Signaling; s31866), acetyl-coenzyme A acetyltransferase 1 (Cell Signaling; s44276), carnitine palmitoyltransferase 1a (Cell Signaling; s97361), glutamate dehydrogenase (Cell Signaling; s12793), vinculin (Cell Signaling; s13901), βactin (Cell Signaling; s4970). After washing, membranes were incubated 1h at room temperature with HRP-linked goat anti-rabbit antibody (Cell Signaling; s7074). Bands were detected using enhanced chemiluminescence (ECL) and their densities quantified using ImageLab software (Bio-rad). Each band was delimited manually, and densities were normalized by dividing by the density of controls [38] (link).
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2

Striatal Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Striatum samples were homogenized by sonication in SDS buffer (10%). After determining the dynamic range of detection for each antibody, 20 μg of total protein were resolved in 4-20% SDS-PAGE and then transferred onto polyvinylidene difluoride (PVDF) membranes using the Trans-Blot Turbo transfer system (Bio-rad). Briefly, membranes were incubated overnight at 4°C with primary antibodies of interest anti-: TH (Cell Signaling; s58844), mTOR (Cell Signaling; s2983), p-mTOR (Cell Signaling; s5536), iNOS (Cell Signaling; s13120), MPC1 (Cell Signaling; s14462), pyruvate dehydrogenase (Cell Signaling; s3205), p-pyruvate dehydrogenase (Cell Signaling; s31866), acetyl-coenzyme A acetyltransferase 1 (Cell Signaling; s44276), carnitine palmitoyltransferase 1a (Cell Signaling; s97361), glutamate dehydrogenase (Cell Signaling; s12793), vinculin (Cell Signaling; s13901), βactin (Cell Signaling; s4970). After washing, membranes were incubated 1h at room temperature with HRP-linked goat anti-rabbit antibody (Cell Signaling; s7074). Bands were detected using enhanced chemiluminescence (ECL) and their densities quantified using ImageLab software (Bio-rad). Each band was delimited manually, and densities were normalized by dividing by the density of controls [38] (link).
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