Polymicrobial Sepsis Induced Monocyte Activation

Animal experiments were approved by the IACUC of BIDMC and done as per NIH guidelines. C57BL/6 wild-type (WT) mice were from Charles River Laboratories (Wilmington, MA). Panx1 knockout (panx1 KO) mice were from The Knockout Mouse Project Center for Comparative Medicine (University of California, Davis, CA) and backcrossed into a C57BL/6 background. Polymicrobial sepsis was induced by cecal ligation and puncture (CLP) as described (19 (link)). In some experiments, mice received i.p. injections of live E. coli (10⁸) and remaining bacteria in the peritoneal cavity were determined after 2 h (19 (link)). Monocyte activation was assessed by flow cytometry using anti-CD11b and anti-Ly6C antibodies. Briefly, blood was collected by cardiac puncture, red blood cells lysed, leukocytes treated with Fc blocker (BD Biosciences), and labeled with PE-anti- Gr1 (clone: RB6–8C5), PerCP-anti-Ly6C (clone: HK1.4), and APC-anti-CD11b (clone: M1/70) antibodies (Thermo Fisher Scientific). Monocytes were identified by gating on Gr1^-CD11b⁺Ly6C⁺ cells.

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.

Lee A.H., Ledderose C., Xiaoou L., Slubowski C.J., Sueyoshi K., Staudenmaier L., Bao Y., Zhang J, & Junger W.G. (2018). ATP release is required for Toll-like receptor-induced monocyte/macrophage activation, NLRP3 inflammasome signaling, IL-1β production, and the host immune response to infection. Critical care medicine, 46(12), e1183-e1189.

Publication 2018

C57BL/6 wild-type (WT) mice Fc blocker APC-anti-CD11b

Anti antibodies Antibodies Bacteria Blood C57bl mice Cardiac Cd11b Cecal Cells Clone E coli Flow cytometry Iacuc Knockout mouse Leukocytes Ligation Medicine Mice Monocytes Peritoneal cavity Puncture Red blood cells River Sepsis

Corresponding Organization :

Other organizations : Beth Israel Deaconess Medical Center, Harvard University, Sichuan University, Ludwig Boltzmann Institute for Experimental and Clinical Traumatology

Top 5 similar protocols

Variable analysis

independent variables

Mouse genotype (C57BL/6 wild-type vs. Panx1 knockout)
Sepsis induction (cecal ligation and puncture, or intraperitoneal injection of E. coli)

dependent variables

Monocyte activation (assessed by flow cytometry using anti-CD11b and anti-Ly6C antibodies)

control variables

Mouse strain (C57BL/6 background)

controls

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.

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?

Revolutionizing how scientists
search and build protocols!