Measuring Skeletal Muscle Fatty Acid Oxidation

Soleus and gastrocnemius muscle of overnight-fasted mice were assayed as described (28 (link)) with modifications (29 (link)). Muscle was homogenized in 10 mmol/L Tris (pH 7.2), 300 mmol/L sucrose, and 2 mmol/L EDTA, injected by syringe into a sealed beaker to be incubated at 30°C for 45 min in the presence of 0.2 mmol/L of [1-¹⁴C]palmitate (0.5 µCi/mL) and 2 mmol/L ATP in incubation buffer (100 mmol/L sucrose, 10 mmol/L Tris-HCl, 5 mmol/L potassium phosphate, 80 mmol/L KCl, 1 mmol/L MgCl₂, 2 mmol/L l-carnitine, 0.1 mmol/L malic acid, 0.05 mmol/L CoA, 1 mmol/L dithiothreitol, 0.2 mmol/L EDTA, and 0.5% BSA, pH 7.4). The reaction was terminated with glacial acetic acid, and trapped CO₂ radioactivity was measured by liquid scintillation in CytoScint (MP Biomedicals).

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Al-Share Q.Y., DeAngelis A.M., Lester S.G., Bowman T.A., Ramakrishnan S.K., Abdallah S.L., Russo L., Patel P.R., Kaw M.K., Raphael C.K., Kim A.J., Heinrich G., Lee A.D., Kim J.K., Kulkarni R.N., Philbrick W.M, & Najjar S.M. (2015). Forced Hepatic Overexpression of CEACAM1 Curtails Diet-Induced Insulin Resistance. Diabetes, 64(8), 2780-2790.

Publication 2015

CytoScint

Buffer Carnitine Dithiothreitol Edta Gastrocnemius muscle Glacial acetic acid Malic acid Mgcl2 Mice Muscle Palmitate Potassium phosphate Radioactivity Soleus Sucrose Syringe Tris

Corresponding Organization : University of Toledo

Other organizations : University of Massachusetts Chan Medical School, Harvard University, Joslin Diabetes Center, Yale University

Top 5 similar protocols

Protocol cited in 1 other protocol

Variable analysis

independent variables

Fasting state (overnight-fasted)

dependent variables

Palmitate oxidation rate in soleus and gastrocnemius muscles

control variables

Incubation temperature (30°C)
Incubation duration (45 min)
Palmitate concentration (0.2 mmol/L)
Palmitate radioactivity (0.5 µCi/mL)
ATP concentration (2 mmol/L)
Composition of incubation buffer (100 mmol/L sucrose, 10 mmol/L Tris-HCl, 5 mmol/L potassium phosphate, 80 mmol/L KCl, 1 mmol/L MgCl2, 2 mmol/L L-carnitine, 0.1 mmol/L malic acid, 0.05 mmol/L CoA, 1 mmol/L dithiothreitol, 0.2 mmol/L EDTA, 0.5% BSA, pH 7.4)
Muscle homogenization buffer (10 mmol/L Tris, pH 7.2, 300 mmol/L sucrose, 2 mmol/L EDTA)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!