SDS-PAGE Protein Analysis Protocol

Cells were harvested in sample buffer (0.1 M Tris [pH 6.8], 4% SDS, 4 mM EDTA, 286 mM 2-mercaptoethanol, 3.2 M glycerol, 0.05% bromophenol blue), and proteins were analyzed as previously described (8 (link)). Commercial antibodies used in this study are as follows: MNV Propol (9 (link)) ATG16L1 and LC3B (Sigma-Aldrich); GBP1-5 and Actin-HRP (Santa Cruz Biotechnology); HA (the Frank W. Fitch Monoclonal Antibody Facility, The University of Chicago); Flag-M2 (Thermo Fischer); WIPI2 (Abcam); HRP Goat anti-mouse and HRP Donkey anti-rabbit (Bio Legend); and HRP Donkey anti-goat (Jackson Immuno Research).

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Kurhade C., Kang S., Biering S.B., Hwang S, & Randall G. (2023). CAPRIN1 Is Required for Control of Viral Replication Complexes by Interferon Gamma. mBio, 14(3), e00172-23.

Publication 2023

WIPI2 HRP Donkey anti-goat

Actin Antibodies Bromophenol blue Buffer Cells Donkey Edta Glycerol Goat Mercaptoethanol Monoclonal antibody Mouse Propol Proteins Rabbit Tris

Corresponding Organization :

Other organizations : University of Chicago, University of California, Berkeley

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein levels

control variables

Sample buffer composition (0.1 M Tris [pH 6.8], 4% SDS, 4 mM EDTA, 286 mM 2-mercaptoethanol, 3.2 M glycerol, 0.05% bromophenol blue)
Experimental methods (proteins were analyzed as previously described (8 (link)))

controls

Positive controls: None explicitly mentioned
Negative controls: None explicitly mentioned

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