Measuring Protein Synthesis in Neuro 2a Cells

Protein synthesis was assesed by L-[ring-3,5-³H]-Tyrosine (Perkin Elmer, Waltham MA) as previously described by [19 (link)]. L-[ring-3,5-³H]-Tyrosine was added for 1 h to Neuro 2a cells that have preincubated for 2h in the absence (Control cells) or presence of 25 μM HMB (HMB cells) in DMEM with 10% FBS and 0.8 mM L-Tyrosine. Radioactivity in dissolved precipitates was counted using a scintillation counter (Beckman Coulter). Data was computed as dpm/μg of proteins.

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Salto R., Vílchez J.D., Girón M.D., Cabrera E., Campos N., Manzano M., Rueda R, & López-Pedrosa J.M. (2015). β-Hydroxy-β-Methylbutyrate (HMB) Promotes Neurite Outgrowth in Neuro2a Cells. PLoS ONE, 10(8), e0135614.

Publication 2015

L-[ring-3,5-3H]-Tyrosine scintillation counter

Cells Protein synthesis Proteins Radioactivity Scintillation counter Tyrosine

Corresponding Organization :

Other organizations : Universidad de Granada, Abbott (Spain)

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Variable analysis

independent variables

Presence or absence of 25 μM HMB

dependent variables

Radioactivity in dissolved precipitates (dpm/μg of proteins)

control variables

DMEM with 10% FBS and 0.8 mM L-Tyrosine
Incubation time of 1 h with L-[ring-3,5-³H]-Tyrosine

controls

Negative control: Neuro 2a cells incubated in the absence of 25 μM HMB (Control cells)
Positive control: Neuro 2a cells incubated in the presence of 25 μM HMB (HMB cells)

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