LPS-Induced NLRP3 Inflammasome Activation
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 :
Other organizations : Trinity College Dublin, University of Queensland, National Human Genome Research Institute, National Institutes of Health, University of Michigan–Ann Arbor, University Hospital Bonn, University of Bonn, Walter and Eliza Hall Institute of Medical Research
Protocol cited in 13 other protocols
Variable analysis
- Stimulation with 10 ng/ml LPS for 3 h
- Addition of 10 µM nigericin for 1 h
- Addition of indicated inhibitors or controls for 30 min
- Cell lysate analysis by Western blotting
- BMDM cells seeded at 10^6/ml in 6 well plates
- Replacement of medium the following day
- Lysis buffer composition (20 mM Hepes-KOH, pH 7.5, 150 mM KCL, 1% NP-40, 0.1 mM PMSF, 1 mg/ml leupeptin, 11.5 mg/ml aprotinin, 1 mM sodium orthovanadate)
- Cross-linking with 2 mM disuccinimydyl suberate (DSS) for 30 min at room temperature
- Negative controls: Indicated inhibitors
- Positive control: Not explicitly mentioned
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!