2 glycerophosphate

Manufactured by Merck Group

2-glycerophosphate is a chemical compound that serves as a laboratory reagent. It is commonly used as a buffering agent in cell culture media and biochemical assays. The core function of 2-glycerophosphate is to maintain a stable pH environment for various experimental applications.

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Lab products found in correlation

Infinity triglycerides reagent, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Bovine intestinal alkaline phosphatase, by Merck Group (1 mentions) Irdye secondary antibody, by LI COR (1 mentions) Odyssey clx infrared imaging system, by LI COR (1 mentions) Benzonase nuclease, by Merck Group (1 mentions) Odyssey blocking buffer, by LI COR (1 mentions) Sodium orthovanadate, by Merck Group (1 mentions) Sodium fluoride, by Merck Group (1 mentions) Protease inhibitor cocktail tablet, by Roche (1 mentions)

3 protocols using 2 glycerophosphate

Quantifying Hepatic Triglyceride Levels

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Triglyceride content within mouse liver was analyzed as previously described [67 (link)]. In brief, liver was homogenized in NETN lysis buffer supplemented with 5 mM NaF (Sigma-Aldrich, St. Louis, MO), 50 mM 2-glycerophosphate (Sigma-Aldrich), and protease inhibitor cocktail (Roche, Nutley, NJ). Lipid content was determined using Infinity Triglycerides Reagent (Thermo Fisher Scientific, Waltham, MA) according to the manufacturer's protocol.

Stout M.B., Tchkonia T., Pirtskhalava T., Palmer A.K., List E.O., Berryman D.E., Lubbers E.R., Escande C., Spong A., Masternak M.M., Oberg A.L., LeBrasseur N.K., Miller R.A., Kopchick J.J., Bartke A, & Kirkland J.L. (2014). Growth hormone action predicts age-related white adipose tissue dysfunction and senescent cell burden in mice. Aging (Albany NY), 6(7), 575-586.

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Dephosphorylation of Yeast Proteins

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Bovine intestinal alkaline phosphatase (Sigma-Aldrich) was used for in vitro protein dephosphorylation reactions following the manufacturer's instructions. In brief, whole-cell extracts of ∼6 × 10⁷ yeast cells precipitated with trichloroacetic acid (TCA, Sigma-Aldrich) were dissolved in 150 μl of phosphatase reaction buffer (5 mM Tris–HCl pH 8.0, 10 mM NaCl, 1 mM MgCl₂ and 0.1 mM dithiothreitol), and then 3–4 μl of 2 M Tris–Base was added to adjust the pH to 7.9. Dephosphorylation reactions were carried out by mixing 37.5 μl of whole-cell extracts with 100 U of Bovine intestinal alkaline phosphatase, followed by incubation for 4 h at 30°C. In the negative control experiments, the phosphatase inhibitor 2-glycerophosphate (Sigma-Aldrich) was added to a final concentration of 16 μM. Dephosphorylation reactions were stopped by the addition of 7% TCA for the subsequent precipitation. The pellet was resuspended in protein sample buffer and then incubated for 10 min at 65°C before analyses using SDS polyacrylamide gel electrophoresis.

Woo T.T., Chuang C.N., Higashide M., Shinohara A, & Wang T.F. (2020). Dual roles of yeast Rad51 N-terminal domain in repairing DNA double-strand breaks. Nucleic Acids Research, 48(15), 8474-8489.

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Western Blot Analysis of Protein Lysates

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Cells were lysed in either RIPA lysis buffer or NP-40 lysis buffer supplemented with 1 mM sodium orthovanadate (Sigma Aldrich), 1 mM sodium fluoride (Sigma Aldrich), 40 mM 2-glycerophosphate (Sigma Aldrich), benzonase nuclease (Sigma Aldrich), and protease inhibitor cocktail tablets (Roche). Protein was quantified using bicinchoninic acid (BCA) assay (Pierce). Lysate was run on a gradient 4–20% SDS-PAGE gel (Biorad) and then transferred to an Immobilon F polyvinylidene difluoride membrane (Millipore). The membranes were blocked using Odyssey blocking buffer (Li-COR) for at least 1 h and then probed with the indicated primary antibodies overnight at 4 °C with rocking. Blots were washed 3 times with Tris-buffered saline with Tween 20 (TBST) and then probed with IRDye secondary antibody (Li-COR) diluted in Li-COR buffer for at least 1 h at room temperature. Blots were washed 3 times in TBST and then scanned with an Odyssey CLx infrared imaging system (Li-COR) for fluorescent blots.

Vogt D., Zaver S., Ranjan A., DiMaio T., Gounder A.P., Smith J.G, & Lagunoff M. (2019). STING is dispensable during KSHV infection of primary endothelial cells. Virology, 540, 150-159.

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