For protein isolation and western blotting, cells were washed with 1X PBS and lysed in radio-immunoprecipitation assay (RIPA) lysis buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium Deoxycholate, 0.1% SDS supplemented with protease inhibitor tablets). Protein quantification and western blotting were carried out as previously described (36 (link)). Antibody against MUC1 has been previously described (51 (link)). Antibodies against GRP78, CHOP, NRF2 and O-GlcNAc were from Cell Signaling Technology. CDA antibody was from Santa Cruz Technology (Dallas, TX USA).
Protocol detail
Protein Isolation and Western Blotting
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.
Antibodies
Antibody
Assay
Buffer
Cells
Chop
Grp78
Immunoprecipitation
Isolation
Muc1
Nacl
Np 40
Nrf2
Protease inhibitor
Protein
Sodium deoxycholate
Tris
Corresponding Organization :
Other organizations : University of Nebraska Medical Center
Top 5 similar protocols
Variable analysis
independent variables
- Protein isolation and western blotting method
- Protein levels of MUC1, GRP78, CHOP, NRF2, O-GlcNAc, and CDA
- Lysis buffer composition (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium Deoxycholate, 0.1% SDS supplemented with protease inhibitor tablets)
- Protein quantification and western blotting procedure (as previously described in 36)
- Positive control: None specified
- Negative control: None specified
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
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!