For in vivo experiments, EROS knockout mice were injected with 250 ng/mL of LPS (Invivogen). Then 2 hr later, 30 mM of ATP or PBS (for control) was injected intraperitoneally. Mice were culled 2 hr later, and the peritoneal cavity washed. Supernatants and serum were analysed for the presence of the cytokines indicated in the figures using multiplex kits from Meso Scale Discovery (MD, USA) and were conducted at the Core Biochemical Assay Laboratory (Cambridge University Hospital, UK).
Multiplex kits
Manufactured by Mesoscale
Sourced in United States
Multiplex kits are laboratory tools designed to analyze multiple analytes or targets simultaneously within a single sample. They enable the detection and quantification of various biomolecules, such as proteins or nucleic acids, in a streamlined and efficient manner.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using multiplex kits
1
EROS Knockout Mice LPS-ATP Cytokine Response
2
Cytokine response to Salmonella infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BM-derived macrophages were prepared as described above and 2 × 105 cells in 100 µl of complete RPMI medium were plated in flat bottomed 96-well tissue culture plates (Cellstar). They were cultured alone or in the presence of live S. Typhimurium M525 (phoN::tetC) at 50 CFU per cell. Supernatants from stimulation of BM-derived macrophages were harvested at 18 h. Alternatively, serum was harvested after euthanasia of mice and subsequent cardiac puncture. Supernatants and serum were analyzed for the presence of the cytokines indicated in the figure using multiplex kits from Mesoscale discovery. Electrochemical luminescence was used as the detection system, and samples were analyzed according to the manufacturer’s protocol and read on a Sector 6000 plate reader. Results were calculated using the Mesoscale Workbench software package.
3
Multiplex Inflammatory Cytokine Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used an electrochemiluminescence assay to detect a proinflammatory panel by multiplex kits obtained from Meso Scale Discovery (Rockville, MD). The kits included individual assay for quantification of IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, and TNF-α, and the chemokine CXCL1. We followed the manufacturer’s protocol for evaluation of plasma samples. We read the results on MSD multi-array imaging platform. We carried out the data analyses using the MSD discovery workbench software.
4
Secreted Proteins in M.tb-Macrophage Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Supernatants from culture wells of uninfected and infected AMs from 28 donors at 2, 24, and 72h were analyzed for secreted proteins relevant to M.tb-macrophage interactions using multiplex kits from Meso Scale Discovery (MSD) (Rockville, MD). The supernatants were collected at 2h and replaced with fresh medium, followed by continuing incubation until 24 or 72h. Twenty-seven secreted proteins were measured: 4 inflammatory mediators (TNF-α, IL-6, IL-1β, IL-10; V-PLEX kit), 6 cytokines [CSF2 (GM-CSF), IL-15, IL-16, IL-1α, IL-7, VEGFA] and 12 chemokines [IP-10 (CXCL10), MCP-1 (CCL2), MCP-4 (CCL13), MDC (CCL22), IL-8, TARC (CCL17), MIP-1α (CCL3; U-PLEX (customized multiplexing)), MIP-1β (CCL4), ENA-78 (CXCL5), IL-18, MIP-3α (CCL20), MIP-3β (CCL19)], and 5 matrix metallo-proteinases (MMP-1, MMP-2, MMP-3, MMP-9, MMP-10; MMP 3-PLEX and MMP 2-PLEX).
5
Biomarker Profiling of Inflammatory and Hemostatic Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Approximately 50 mL of venous blood was obtained from each subject. A portion of fresh blood was sent to a clinical laboratory (Lab Corp., Burlington, NC, USA) for analysis of differential blood cell counts, as well as quantification of blood lipids. The remaining blood samples were stored at −80 °C prior to analysis.
Commercially available multiplex kits (Meso Scale Diagnostics, Gaithersburg, MD) were used to quantify levels of C-reactive protein (CRP), serum amyloid A (SAA), soluble intercellular adhesion molecule (sICAM), soluble vascular adhesion molecule (sVCAM), interleukin 1-beta (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α) (SECTOR® Imager 2400, Meso Scale Diagnostics). All other assays (D-dimer, tissue plasminogen factor (tPA), von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1), and plasminogen) were measured using MesoScale multi-array plates as per manufacturers’ instructions.
Commercially available multiplex kits (Meso Scale Diagnostics, Gaithersburg, MD) were used to quantify levels of C-reactive protein (CRP), serum amyloid A (SAA), soluble intercellular adhesion molecule (sICAM), soluble vascular adhesion molecule (sVCAM), interleukin 1-beta (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α) (SECTOR® Imager 2400, Meso Scale Diagnostics). All other assays (D-dimer, tissue plasminogen factor (tPA), von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1), and plasminogen) were measured using MesoScale multi-array plates as per manufacturers’ instructions.
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!