Protocol detail

Neocortex Lysate Western Blot Analysis

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Analysis of neocortex lysates by western blot was performed as described^{18 (link)}. The following primary antibodies were used: anti-eIF4EBP1 (1:1,000, rabbit, Abcam, ab32024, RRID:AB_2097990), anti-phospho-eIF4EBP1 Thr37/46 (1:1,000, rabbit, Cell Signaling Technology, 2855, RRID:AB_560835), and anti-Gapdh (1:1,000, mouse, Millipore, MAB374, RRID:AB_2107445). All HRP secondary antibodies were used at a dilution of 1:2,500: HRP anti-mouse heavy chain (goat, Abcam, ab97245; RRID:AB_10680049) and HRP anti-rabbit heavy chain (goat, Cell Signaling Technology, 7074S, RRID:AB_2099233).

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Harnett D., Ambrozkiewicz M.C., Zinnall U., Rusanova A., Borisova E., Drescher A.N., Couce-Iglesias M., Villamil G., Dannenberg R., Imami K., Münster-Wandowski A., Fauler B., Mielke T., Selbach M., Landthaler M., Spahn C.M., Tarabykin V., Ohler U, & Kraushar M.L. (2022). A critical period of translational control during brain development at codon resolution. Nature Structural & Molecular Biology, 29(12), 1277-1290.

Publication 2022

ab32024 MAB374 ab97245

Antibodies Dilution Gapdh Goat Mouse Neocortex Rabbit Western blot

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Variable analysis

independent variables

Primary antibodies used for western blot analysis

dependent variables

Protein expression and phosphorylation levels detected by western blot

control variables

Sample preparation and western blot analysis procedure, as described in the referenced study (link provided)

controls

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

Annotations

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