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3h l phenylalanine

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[3H]-L-phenylalanine is a radioactively labeled amino acid used in various research applications. It is a synthetic compound that incorporates tritium (3H) into the L-phenylalanine molecule. This product can be used as a tracer or precursor in studies involving protein synthesis, amino acid metabolism, and other related research areas.

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Lab products found in correlation

Beckman ls 6500 multi purpose scintillation counter, by Beckman Coulter (2 mentions) 2 aminobicyclo 2 2 1 heptane 2 carboxylic acid bch, by Merck Group (2 mentions) Hank s balanced salt solution, by Thermo Fisher Scientific (2 mentions) Analytical grade reagents, by Merck Group (1 mentions) 14c sucrose, by PerkinElmer (1 mentions) 3h mpp, by PerkinElmer (1 mentions) Complete mini cocktail, by Roche (1 mentions) Mass spectrometry grade rlys c, by Promega (1 mentions) Tetraethyl ammonium tea, by Merck Group (1 mentions) Sodium dodecyl sulfate (sds), by Merck Group (1 mentions) Sequencing grade modified trypsin, by Promega (1 mentions) Proteasemax surfactant, by Promega (1 mentions) 3h l kynurenine, by PerkinElmer (1 mentions) Mmessage mmachine sp6 kit, by Thermo Fisher Scientific (1 mentions)

4 protocols using 3h l phenylalanine

1

Radiotracer Quantification Protocol

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[14C]-Sucrose (580–600 mCi/mmol), [3H]-L-phenylalanine (128 Ci/mmol) and [3H]-MPP+ (70–84 Ci/mmol) were purchased from Perkin Elmer (Paris, France). Protease Inhibitor Cocktail Complete Mini® was purchased from Roche (Basel, Switzerland). Protease MAX surfactant, mass spectrometry grade rLys-C and sequencing grade modified trypsin were acquired from Promega (Charbonnières-les-Bains, France). RIPA buffer was prepared employing analytical grade reagents from Sigma Aldrich (Saint Quentin Fallavier, France): 50 mmol L−1 Tris (pH 8.0), 150 mmol L−1 NaCl, 1% (V/V) Triton X-100, 0.1% (V/V) sodium dodecyl sulfate (SDS) and 0.5% (W/V) sodium deoxycholate in high purity water. Standard peptides for protein quantification were provided by Bertin Pharma (Orleans, France). Tetraethylammonium (TEA) was purchased from Sigma Aldrich. All other chemicals were of analytical grade.
Chaves C., Campanelli F., Chapy H., Gomez-Zepeda D., Glacial F., Smirnova M., Taghi M., Pallud J., Perrière N., Declèves X., Menet M.C, & Cisternino S. (2020). An Interspecies Molecular and Functional Study of Organic Cation Transporters at the Blood-Brain Barrier: From Rodents to Humans. Pharmaceutics, 12(4), 308.
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2

Amino Acid Uptake Assay in Cells

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Between 0.5 and 1×106 cells were resuspended in 0.4 ml Hank’s balanced-salt solution (Life Technologies) containing 3HL-phenylalanine (0.5 μCi/ml) (Perkin Elmer) with a final extracellular L-leucine concentration of 5 μM. Where indicated, 10 mM BCH (2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, Sigma) was added to the uptake medium. Amino acid uptake was assayed for 4 minutes with samples layered over 0.5 ml of a mixture of silicone oil (Dow Corning 550 (BDH silicone products); specific density, 1.07 g/ml) and dibutyl phthalate (Fluka) at a ratio of 1:1. Cells were pelleted below the oil, the uptake media above the oil layer aspirated. Following 3 washes, the silicon oil–dibutyl phthalate mixture was aspirated, and the cell pellet underneath was resuspended in 200 μl NaOH (500 mM). β-radioactivity was measured by liquid scintillation counting in a Beckman LS 6500 Multi-Purpose Scintillation Counter (Beckman Coulter). Each data point was performed in triplicate.
Pallett L.J., Gill U.S., Quaglia A., Sinclair L.V., Jover-Cobos M., Schurich A., Singh K.P., Thomas N., Das A., Chen A., Fusai G., Bertoletti A., Cantrell D.A., Kennedy P.T., Davies N.A., Haniffa M, & Maini M.K. (2015). Metabolic regulation of hepatitis B immunopathology by myeloid-derived suppressor cells. Nature medicine, 21(6), 591-600.
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3

Radiolabeled Amino Acid Uptake in Oocytes

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All capped RNA (cRNA) was synthesized using the mMessage mMachine SP6 kit (Ambion) and injected into oocytes by Ecocyte Bio Science LLC. Oocytes were incubated at 18°C in modified Barth's solution with gentamicin that was replenished every 24 h followed by three washes and preincubation for at least 5 min at 25°C in uptake buffer. For uptake assays, sodium-free modified Barth's solution containing labeled [3H]-L-phenylalanine or [3H]-L-kynurenine (PerkinElmer) was used. Oocytes were then washed five times in ice-cold uptake buffer and lysed in 0.5 mL of 1% SDS solution for at least 1 h. Radioactivity (disintegrations per minute) was measured using a liquid scintillation counter machine. A detailed procedure is described in Supplemental Material.
Lin L., Lemieux G.A., Enogieru O.J., Giacomini K.M, & Ashrafi K. (2020). Neural production of kynurenic acid in Caenorhabditis elegans requires the AAT-1 transporter. Genes & Development, 34(15-16), 1033-1038.
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4

Amino Acid Uptake Assay in Cells

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Between 0.5 and 1×106 cells were resuspended in 0.4 ml Hank’s balanced-salt solution (Life Technologies) containing 3HL-phenylalanine (0.5 μCi/ml) (Perkin Elmer) with a final extracellular L-leucine concentration of 5 μM. Where indicated, 10 mM BCH (2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, Sigma) was added to the uptake medium. Amino acid uptake was assayed for 4 minutes with samples layered over 0.5 ml of a mixture of silicone oil (Dow Corning 550 (BDH silicone products); specific density, 1.07 g/ml) and dibutyl phthalate (Fluka) at a ratio of 1:1. Cells were pelleted below the oil, the uptake media above the oil layer aspirated. Following 3 washes, the silicon oil–dibutyl phthalate mixture was aspirated, and the cell pellet underneath was resuspended in 200 μl NaOH (500 mM). β-radioactivity was measured by liquid scintillation counting in a Beckman LS 6500 Multi-Purpose Scintillation Counter (Beckman Coulter). Each data point was performed in triplicate.
Pallett L.J., Gill U.S., Quaglia A., Sinclair L.V., Jover-Cobos M., Schurich A., Singh K.P., Thomas N., Das A., Chen A., Fusai G., Bertoletti A., Cantrell D.A., Kennedy P.T., Davies N.A., Haniffa M, & Maini M.K. (2015). Metabolic regulation of hepatitis B immunopathology by myeloid-derived suppressor cells. Nature medicine, 21(6), 591-600.
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