This method was modified from our published study [6 (link)]. In brief, human fibroblasts cells were lysed in immunoprecipitation buffer (Cat# 87788, Thermo scientific) and incubated with primary antibodies overnight at 4°C. Protein G agarose beads (Cat# 15920-010, Life technologies) were added for another 2 hour incubation at 4°C. Proteins were loaded into SDS-PAGE gels, transferred to a PVDF membrane, blotted with primary antibodies and secondary antibodies.
Protocol detail
Immunoprecipitation of Cellular 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.
Agarose
Antibodies
Buffer
Fibroblasts cells
Gels
Human
Immunoprecipitation
Membrane
Protein g
Proteins
Pvdf
Sds page
Corresponding Organization : Vanderbilt University Medical Center
Top 5 similar protocols
Variable analysis
independent variables
- Lysing human fibroblasts cells in immunoprecipitation buffer
- Incubating cell lysates with primary antibodies overnight at 4°C
- Adding Protein G agarose beads and incubating for 2 hours at 4°C
- Proteins loaded into SDS-PAGE gels
- Proteins transferred to a PVDF membrane
- Proteins blotted with primary and secondary antibodies
- Immunoprecipitation buffer (Cat# 87788, Thermo scientific)
- Protein G agarose beads (Cat# 15920-010, Life technologies)
- Incubation temperatures at 4°C
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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!