Proteins were loaded on 12% polyacrylamide gel, with color Prestained Protein Standard (NEB). After migration, proteins were stained with Instant Blue (Abcam). For Western blotting, proteins were transferred onto a nitrocellulose membrane (Amersham Protran, Cytiva) using a Power Blotter apparatus (Invitrogen). Membranes were then saturated in PBS containing 0.1% Tween 20 (PBST) and 4 % skim milk overnight. They were incubated for 2 h with PBST containing a 1:2000 dilution of mouse monoclonal anti-polyHistidine antibody (Sigma-Aldrich) or alternatively with mouse monoclonal anti-MBP antibody (NEB), and then with PBST containing a 1:1000 dilution of Goat anti-mouse IgG Peroxydase conjugated (Invitrogen) for 1 h. Immunodetected proteins were revealed with Pierce ECL Western Blotting Substrate (Thermo), following the manufacturer’s instructions, and chemiluminescence was detected with a ChemiDoc XRS+ system (Bio-Rad).
Power blotter apparatus
Manufactured by Thermo Fisher Scientific
The Power Blotter apparatus is a compact and versatile laboratory instrument designed for efficient protein transfer and blotting applications. It facilitates the electrophoretic transfer of proteins from polyacrylamide gels to membranes, enabling further analysis and detection. The Power Blotter provides consistent and reliable performance for a wide range of blotting needs in research and diagnostic settings.
Automatically generated - may contain errors
Lab products found in correlation
Mouse monoclonal anti polyhistidine antibody, by Merck Group (1 mentions)
Mouse monoclonal anti mbp antibody, by New England Biolabs (1 mentions)
Amersham protran, by Cytiva (1 mentions)
Instantblue, by Abcam (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
C digit, by LI COR (1 mentions)
Chemidoc, by Bio-Rad (1 mentions)
2 protocols using power blotter apparatus
1
Western Blotting of Polyhistidine-Tagged Proteins
2
Western Blotting Protein Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collected cell pellets were lysed in RIPA and separated by electrophoresis on 4–12% NuPAGE Bis-Tris Mini Protein gels (Invitrogen), followed by semi-dry transfer to nitrocellulose membranes using a Power Blotter apparatus (Invitrogen). After blocking in 3% milk in TBST, membranes were sequentially incubated with primary and secondary antibodies. Washed blots were imaged using a ChemiDoc (BioRad) or c-Digit (LiCor) apparatus. Antibodies used for these studies are listed in Supplementary Table 5 .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!