SDS-PAGE and Western Blotting Assay

SDS-PAGE and Western blotting were performed as described [24 (link)]. We used the following polyclonal antibodies: rabbit anti-CspE antibodies [15 (link)]; anti-ribosomal protein antibodies from our lab collection; anti-SecE antibodies were obtained from Dr. Hajime Tokuda; goat anti-rabbit antibodies conjugated to horseradish peroxidase served as secondary antibodies (Jackson Immunoresearch). 6His-tagged proteins were detected by His-probe (Thermo-Fisher™).

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Benhalevy D., Biran I., Bochkareva E.S., Sorek R, & Bibi E. (2017). Evidence for a cytoplasmic pool of ribosome-free mRNAs encoding inner membrane proteins in Escherichia coli. PLoS ONE, 12(8), e0183862.

Publication 2017

goat anti-rabbit antibodies His-probe

Anti antibodies Antibodies Goat Horseradish peroxidase Proteins Rabbit Ribosomal protein Sds page

Corresponding Organization : Weizmann Institute of Science

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of CspE, ribosomal proteins, and SecE

control variables

SDS-PAGE and Western blotting protocols as described in [24]
Polyclonal antibodies used: rabbit anti-CspE, anti-ribosomal protein, anti-SecE, and goat anti-rabbit antibodies conjugated to horseradish peroxidase
6His-tagged proteins detected by His-probe (Thermo-Fisher™)

controls

Positive control: 6His-tagged proteins detected by His-probe
Negative control: Not explicitly mentioned

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