Cells were lysed using RIPA buffer, and lysates were prepared for electrophoresis with NuPAGE™ sample reducing agent (Thermofisher) and NuPAGE™ SDS loading buffer (Thermofisher). Samples were heated to 95°C for 5 minutes. A total of 25 µg of protein was resolved with SDS-PAGE using 3-8% tris-acetate gels for blots probing polyubiquitin or 4-12% bis-tris gels for all the others. Proteins were transferred to nitrocellulose membranes using Thermo Scientific™ Pierce™ G2 Fast blot. Membranes were incubated in primary antibody diluted in PBST with 2% BSA overnight at 4°C, washed, and incubated with the appropriate secondary antibody diluted in PBST with 5% skimmed milk. Blots were developed using Clarity Western ECL substrate (Bio-Rad) and imaged using ChemiDoc MP Imager (Biorad). All images were processed in ImageLab v 5.2.1 (Biorad).
Pierce g2 fast blot
Manufactured by Thermo Fisher Scientific
The Pierce™ G2 Fast blot is a rapid and efficient protein transfer system designed for Western blotting. It enables fast and reliable transfer of proteins from polyacrylamide gels to membranes, facilitating downstream detection and analysis.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pierce g2 fast blot
1
Western Blot Protocol for Protein Analysis
2
Western Blot Analysis of Polyubiquitin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed using RIPA buffer, and lysates were prepared for electrophoresis with NuPAGE™ sample reducing agent (Thermofisher) and NuPAGE™ SDS loading buffer (Thermofisher). Samples were heated to 95 °C for 5 min. A total of 25 µg of protein was resolved with SDS-PAGE using 3-8% tris-acetate gels for blots probing polyubiquitin or 4-12% bis-tris gels for all the others. Proteins were transferred to nitrocellulose membranes using Thermo Scientific™ Pierce™ G2 Fast blot. Membranes were incubated in primary antibody diluted in PBST with 2% BSA overnight at 4 °C, washed, and incubated with the appropriate secondary antibody diluted in PBST with 5% skimmed milk. Blots were developed using Clarity Western ECL substrate (Bio-Rad) and imaged using ChemiDoc MP Imager (Biorad). Images were acquired with ImageLab v 5.2.1 (Biorad) using the built-in protocol "Chemi Hi-sensitivity" via autoexposure or manual exposure. Global contrast and gamma adjustments were done on the images. All antibodies used are in Supplementary Table 3.
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!