1 2 14c acetate

Manufactured by PerkinElmer

Sourced in United States, Italy

The [1,2-14C]acetate is a radioactive organic compound labeled with the carbon-14 isotope. It is commonly used as a research tool in various scientific applications, particularly in the fields of biochemistry, molecular biology, and tracer studies. The core function of this product is to serve as a labeled precursor for the incorporation of radioactive carbon into organic molecules or metabolic pathways, enabling researchers to study and track specific chemical reactions and metabolic processes.

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Lab products found in correlation

Aminoglutethimide, by Merck Group (1 mentions) Cellstripper, by Corning (1 mentions) 1 14c oleate, by PerkinElmer (1 mentions) 14c acetate, by PerkinElmer (1 mentions)

Spelling variants of this product

[14C]-acetate (22 citations) [1-14C]-acetate (19 citations) Sodium [1,2-14C] acetate (4 citations) [2-14C]acetate (2 citations)

3 protocols using 1 2 14c acetate

Measuring Cholesterol Biosynthesis in Y-1 Cells

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De novo cholesterogenic activity was measured as described previously^{47 (link),48 (link)}. Y-1 cells were treated with siAd4BP/SF-1 (#1635) or siControl for 48 h, and then incubated for 1 h at 37 °C in a serum-free medium containing [1,2-¹⁴C]acetate (PerkinElmer, Boston, MA, USA), 50 μM aminoglutethimide (Sigma-Aldrich) and 2 μg/ml 58-035 (ACAT2; acyl-CoA cholesterol acyltransferase inhibitor) (Sigma-Aldrich). Total lipid fraction containing cholesterol was extracted with chloroform/methanol (2:1, v/v), and then separated by thin-layer chromatography on a silica gel using benzene-ethylacetate (2:3, v/v) as a solvent. The radioactivities of the spots for free and esterified cholesterol, visualized with iodine vapor, were determined by liquid scintillation counting. [1,2-¹⁴C]acetate incorporation into cholesterol was expressed as cpm/mg protein/h.

Baba T., Otake H., Inoue M., Sato T., Ishihara Y., Moon J.Y., Tsuchiya M., Miyabayashi K., Ogawa H., Shima Y., Wang L., Sato R., Yamazaki T., Suyama M., Nomura M., Choi M.H., Ohkawa Y, & Morohashi K.I. (2018). Ad4BP/SF-1 regulates cholesterol synthesis to boost the production of steroids. Communications Biology, 1, 18.

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Lipid Synthesis Analysis in Insulin-Stimulated Hepatocytes

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Isolated primary hepatocytes of WT, Lrp6^mut/mut, Ldlr^−/− and Ldlr^−/−/Lrp6^mut/mut mice were serum starved overnight, followed by insulin stimulation (100 nM) for 4 h. Insulin stimulated primary hepatocytes were incubated with 25.0 nmol of [1,2-¹⁴C]-acetate (Perkin Elmer), 12.5 nmol [1-¹⁴C]-plamitate (Perkin Elmer), and 12.5 nmol [1-¹⁴C]-oleate (Perkin Elmer) for 2h. Cells were washed three times with PBS before harvest in Cellstripper (Cellgro). Total lipid from cells was prepared by the Bligh and Dyer method (Bligh and Dyer, 1959 (link)). Concentrated neutral lipids were dissolved in chloroform: methanol (2:1, v/v) and lipid fractions were separated by thin layer chromatography (TLC, Silica 60 F₂₅₄, Merck) using the organic mobile solvents hexane/diethylether/acetic acid (70:30:1 v/v). Lipids dots were visualized under UV light and radioactivity was determined using a scintillation counter and normalized to protein concentrations.

Go G.W., Srivastava R., Hernandez-Ono A., Gang G., Smith S.B., Booth C.J., Ginsberg H.N, & Mani A. (2014). The Combined Hyperlipidemia Caused by Impaired Wnt-LRP6 signaling is reversed by Wnt3a Rescue. Cell metabolism, 19(2), 209-220.

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Sterol Biosynthesis Tracking in Yeast

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Sterol biosynthesis in whole yeast cells was followed by incorporation of [ 14 C] acetate into non-saponifiable lipids as described previously [30] . Briefly, washed cells (1×10 8 cells) were resuspended in 1ml of fresh media, incubated with 0.1 μCi of [ 14 C]acetate ([1,2-14 C]acetate (50 μCi, 1.85 MBq) from Perkin-Elmer, Italy) in presence of Tween-80 (0.2 mg/ml) and shaken for 3 h at 30°C. Cells were then saponified in 1ml of methanolic KOH (15% w/v) for 30 min at 80°C. Non-saponifiable lipids were extracted with petroleum ether and separated on TLC plates (20×20 cm×0.25 mm) using cyclohexane/ethyl acetate (85:15; v/v). Squalene, oxidosqualene, dioxidosqualene, lanosterol, ketosteroid 1 (a mixture of 4-methylzymosterone and 4methylfecosterone), ketosteroid 2 (a mixture of zymosterone and episterone) and ergosterol were used as references. Radioactivity in separated bands was measured by collecting counts over a 5 min period with a System 200 Imaging Scanner (Hewlett-Packard, Palo Alto, CA, USA).

Cirigliano A., Macone A., Bianchi M.M., Oliaro-Bosso S., Balliano G., Negri R, & Rinaldi T. (2019). Ergosterol reduction impairs mitochondrial DNA maintenance in S. cerevisiae. Biochimica et biophysica acta. Molecular and cell biology of lipids, 1864(3).

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