Immunoblotting Protocol for Protein Analysis

Immunoblotting was performed as previously described^{17 (link)}. Briefly, cells were lysed in RIPA buffer, and protein lysates were resolved on Novex 4–12% or 10–20% Tris-Glycine gels (Invitrogen, Carlsbad, CA), transferred to nitrocellulose (Bio-Rad, Hercules, CA) and then incubated with primary antibodies (see Table S2 for primary antibodies). All immunoblotting data shown is a representative of at least 3 biological replicates.

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Aksoylu I.S., Martin P., Robert F., Szkop K.J., Redmond N.E., Chen S., Beauchamp R.L., Nobeli I., Pelletier J., Larsson O, & Ramesh V. (2023). Translatome analysis of Tuberous Sclerosis Complex-1 patient-derived neural progenitor cells reveal rapamycin-dependent and independent alterations. Research Square.

Publication Preprint 2023

Novex 4–12% –20% Tris-Glycine gels nitrocellulose

Antibodies Biological Buffer Cells Gels Glycine Nitrocellulose Protein Ripa Tris

Corresponding Organization :

Other organizations : Karolinska Institutet, Massachusetts General Hospital, McGill University, Birkbeck, University of London

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Not explicitly mentioned

control variables

Protein lysates were resolved on Novex 4–12% or 10–20% Tris-Glycine gels (Invitrogen, Carlsbad, CA)
Protein lysates were transferred to nitrocellulose (Bio-Rad, Hercules, CA)
Cells were lysed in RIPA buffer

positive controls

Not specified

negative controls

Not specified

Annotations

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