Bone marrow–derived macrophages were prepared as described previously (21 (link)). Cells were stimulated with LPS (500 ng/mL; InvivoGen), Pam3CSK4 (1 μg/mL), gardiquimod (1 μg/mL; InvivoGen), Poly(I:C) (10 μg/mL; InvivoGen), or IFNβ (400 U/mL; PBL Assay Science). For transfection of DNA or 2´3´-cyclic guanosine monophosphate–adenosine monophosphate (cGAMP), 0.25 μg of poly(dA:dT) (InvivoGen) or 1 μg of 2´3´-cGAMP (InvivoGen) was mixed with 0.3 mL of Xfect polymer in Xfect reaction buffer (Clontech Laboratories) for 10 min and then added to BMDMs in Opti-MEM (ThermoFisher Scientific).
Protocol detail
Bone Marrow Macrophage Stimulation Assay
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.
LPS
Pam3CSK4
gardiquimod
Poly(I:C)
IFNβ
poly(dA:dT)
2´3´-cGAMP
Xfect polymer
Xfect reaction buffer
Opti-MEM
Assay
Bone marrow derived macrophages
Buffer
Cells
Cyclic guanosine monophosphate adenosine monophosphate
Gardiquimod
Poly
Poly i c
Polymer
Transfection
Corresponding Organization : St. Jude Children's Research Hospital
Other organizations : University of Tennessee Health Science Center
Top 5 similar protocols
Variable analysis
independent variables
- LPS (500 ng/mL; InvivoGen)
- Pam3CSK4 (1 μg/mL)
- Gardiquimod (1 μg/mL; InvivoGen)
- Poly(I:C) (10 μg/mL; InvivoGen)
- IFNβ (400 U/mL; PBL Assay Science)
- Poly(dA:dT) (0.25 μg; InvivoGen)
- 2´3´-cGAMP (1 μg; InvivoGen)
- Not explicitly mentioned
- Not explicitly mentioned
- Positive controls: Not specified
- Negative controls: Not 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!