Western Blot Analysis of MEIS Proteins
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : University of Chicago
Other organizations : Johns Hopkins University
Variable analysis
- Whole-cell lysates of 100,000 or more cells were used per lane
- Expression levels of MEIS1 and MEIS2 proteins
- Cells were rinsed with cold PBS
- Cells were scraped into RIPA buffer supplemented with protease inhibitors
- Samples were sonicated twice and re-suspended in 4× sample buffer (BioRad; Hercules, CA) supplemented with 10% Beta-mercaptoethanol (Sigma-Aldrich; St. Louis MO)
- The Pierce BCA protein assay kit (Thermo Fisher Scientific; Grand Island, NY) was used to determine protein concentration
- 60 ug of protein was loaded on a 10% SDS-polyacrylamide gel and electrophoresed
- Proteins were transferred to nitrocellulose (LI COR, Odyssey; Lincoln, NE) overnight at 4°C
- The membrane was blocked overnight in 5% non-fat milk in TBS at 4°C
- The secondary antibodies goat anti-mouse IRDye 800 CW or donkey anti-rabbit IRDye 680 from LI-COR Biosciences were used, and images captured using an infrared Odyssey scanner (LI-COR)
- Not specified
- Not specified
Annotations
Based on most similar protocols
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!