GST Pull-Down Assay with Radiolabeled Proteins

GST pull-down assays with in vitro translated
^{35 (link)}S-labeled proteins
were done as described previously ^{14 (link)}. Briefly, GST-fusion proteins were expressed in Escherichia
coli BL21(DE3) and/or SoluBL21 (Amsbio). After pull-down assays, the proteins
were separated by SDS-PAGE and transferred to PVDF membranes and the
radiolabeled proteins were detected in a PharosFX and PharosFX Plus Imager
(BioRad).

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Kumar S., Jain A., Choi S.W., da Silva G.P., Allers L., Mudd M.H., Peters R.S., Anonsen J.H., Rusten T.E., Lazarou M, & Deretic V. (2020). Mammalian Atg8 proteins and autophagy factor IRGM control mTOR and TFEB at a regulatory node critical for response to pathogens. Nature cell biology, 22(8), 973-985.

Publication 2020

SoluBL21 PharosFX PharosFX Plus Imager

Assays Membranes Proteins Pull Pvdf Sds page

Corresponding Organization :

Other organizations : University of New Mexico, University of Oslo, Australian Regenerative Medicine Institute, Monash University

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Variable analysis

independent variables

GST-fusion proteins expressed in Escherichia coli BL21(DE3) and/or SoluBL21 (Amsbio)

dependent variables

Radiolabeled proteins detected in a PharosFX and PharosFX Plus Imager (BioRad)

control variables

Pull-down assays were done as described previously [14]

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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