Proteins were isolated from whole cells in lysis buffer (20mM Tris-HCl [pH 8], 137mM NaCl, 10% glycerol, 1% Triton X-100, and 2mM EDTA) containing Protease Inhibitor Cocktail 78410 (Thermo Fisher Scientific). Proteins were resolved by polyacrylamide gel electrophoresis on 10% sodium dodecyl sulfate gels EC60752 (Thermo Fisher Scientific) and immobilized onto nitrocellulose membranes (GE Healthcare, Chicago, IL, USA) by electroblot (Bio-Rad Laboratories, Hercules, CA, USA). Specific proteins were detected by immunoblotting with the indicated antibodies (STable 1 ); immunoreactive proteins were detected by enhanced chemiluminesence (Thermo Scientific) on a ChemiDoc XRS+ (Bio-Rad Laboratories).
Electroblot
Manufactured by Bio-Rad
Sourced in United States
The Electroblot is a laboratory equipment used for the transfer of proteins or nucleic acids from a gel to a membrane for further analysis. It facilitates the efficient and accurate transfer of these biomolecules from the gel to the membrane, enabling subsequent detection and study.
Automatically generated - may contain errors
Lab products found in correlation
Nitrocellulose membrane, by GE Healthcare (3 mentions)
Chemidoc xr, by Bio-Rad (3 mentions)
Protease inhibitor cocktail 78410, by Thermo Fisher Scientific (2 mentions)
Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Ma5 15739, by Thermo Fisher Scientific (1 mentions)
Tris glycine gel, by Thermo Fisher Scientific (1 mentions)
Pierce coomassie plus bradford assay, by Thermo Fisher Scientific (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions)
3 protocols using electroblot
1
Protein Isolation and Immunoblotting
2
Protein Extraction and Western Blot Analysis
3
Western Blot Protein Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Proteins were isolated from whole cells in lysis buffer (20mM Tris-HCl (pH 8), 137mM NaCl, 10% glycerol, 1% Triton X-100, and 2mM EDTA) containing Protease Inhibitor Cocktail (Catalogue Number 78410) (Thermo Fisher Scientific), 1 mM Na3VO4, and 1 mM PMSF (Sigma). Proteins were quantified via Pierce Coomassie Plus (Bradford) Assay (Thermo Fisher Scientific), and 25 µg of protein per sample were resolved by polyacrylamide gel electrophoresis on 10% Tris-Glycine gels (Catalogue Number XP00105BOX) (Thermo Fisher Scientific) and immobilized onto nitrocellulose membranes (GE Healthcare, Chicago, IL, USA) by electroblot (Bio-Rad Laboratories, Hercules, CA, USA). Membranes were blocked with 5% w/v BSA in TBS and 0.1% v/v Tween20 (Gold Bio, St Louis, MO, USA) for one hour at room temperature and incubated with primary antibodies overnight (Supplementary Table 1); immunoreactive proteins were detected by enhanced chemiluminescence (Thermo Scientific) on a ChemiDoc XRS+ (Bio-Rad Laboratories). Densitometry analysis in Figure 4B was performed using ImageJ (NIH, Bethesda, MD, USA).
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