- Other Names
- Mouse Macrophage Panel, Macrophage, Multiplex, flow cytometry, LEGENDplex, CXCL1 (KC), TGF-β1, IL-18, IL-23, CCL22 (MDC), IL-10, IL-12p70, IL-6, TNF-α, G-CSF, CCL17 (TARC), IL-12p40, IL-1β
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Macrophages are produced by differentiation of monocyte in response to an infection or tissue damage. Their primary function is to recognize, engulf, and destroy target cells including pathogens, dying or dead cells, and cellular debris. Like dendritic cells, macrophages are also professional antigen presenting cells that play a crucial role in initiating an immune response. Macrophages secrete an array of cytokines which aid in host defense, tissue repair, and immunoregulation. Macrophages can be divided into multiple subtypes based on different functions. Inflammation-encouraging M1 macrophages produce proinflammatory cytokines, such as CXCL1 (KC), IL-18, IL-23, IL-12p70, IL-6, TNF-α, IL-12p40, and IL-1β. Anti-inflammatory and tissue-repairing M2 macrophages decrease immune reactions and promote tissue repair by releasing a different set of factors, such as Free Active TGF-β1, CCL22 (MDC), IL-10, IL-6, G-CSF, and CCL17 (TARC).
Due to their important roles in immune responses, macrophages are critically involved in a variety of inflammatory disorders, such as sepsis-related multiple organ dysfunction/failure, microbial infection, acute injuries, neurodegenerative disorders, cancer, cardiovascular, and autoimmune diseases. Measuring the mediators of proinflammation and anti-inflammation produced by macrophages may help not only in understanding the fundamental functions of macrophages, but also in finding the mechanisms of various pathological processes.
The LEGENDplex™ Mouse Macrophage/Microglia Panel (13-plex) is a bead-based multiplex assay panel, using fluorescence-encoded beads suitable for use on various flow cytometers. It allows for simultaneous quantification of 13 key targets involved in monocyte differentiation and macrophage functions such as CXCL1 (KC), Free Active TGF-β1, IL-18, IL-23, CCL22 (MDC), IL-10, IL-12p70, IL-6, TNF-α, G-CSF, CCL17 (TARC), IL-12p40, and IL-1β. This assay panel provides higher detection sensitivity and broader dynamic range than traditional ELISA methods. The panel has been validated for use on cell culture supernatant and serum.
The LEGENDplex™ Mouse Macrophage/Microglia Panel (13-plex) is designed to allow flexible customization within the panel. It can also be divided into subpanels such as:
LEGENDplex™ Mouse M1 Macrophage Panel (8-plex)
LEGENDplex™ Mouse M2 Macrophage Panel (6-plex)
Please visit www.biolegend.com/legendplex for more information on how to mix and match within the panel.
This assay is for research use only.
- Kit Contents
- Setup Beads 1: FITC Beads (1 ml)
- Setup Beads 2: PE Beads (1 ml)
- Setup Beads 3: Raw Beads (2 ml)
- Capture Beads
- Detection Antibody (3.5 ml)
- Standard Cocktail, Lyophilized (1 vial)
- SA-PE (3.5 ml)
- Matrix C, Lyophilized (1 vial)
- Assay Buffer (25 ml)
- Wash Buffer, 20X (25 ml)
- Filter Plate (1 plate)
- Plate Sealers (4 sheets)
- Data Analysis Software Dongle
- Antibody Type
Learn more about LEGENDplex™ at biolegend.com/legendplex.
Download the LEGENDplex™ software here.
- Materials Not Included
- Flow Cytometer
- Pipettes and Tips
- Reagent Reservoirs for Multichannel Pipettes
- Polypropylene Microfuge Tubes, in 96-Tube Rack
- Vortex Mixer
- Aluminum foil
- Absorbent pads
- Plate Shaker
- A Vacuum Filtration Unit and a Vacuum Source (if using filter plates)
- Biology Area
- Immunology, Innate Immunity, Neuroscience, Neuroinflammation
- Molecular Family
- Gene ID
- 21803 View all products for this Gene ID 20299 View all products for this Gene ID 16153 View all products for this Gene ID 16193 View all products for this Gene ID 12985 View all products for this Gene ID 20295 View all products for this Gene ID
- View information about M2 Macrophage on UniProt.org
- After I finish the staining process, how long can I wait before reading my LEGENDplex™ samples?
The samples can be kept overnight at 4°C while being protected from exposure to light and be read the next day. There may be a decrease in signal, but overall, the assay results should not be affected. Storing the samples for extended periods of time is not recommended, as it could lead to further reductions in signal.
- If I don't have a vacuum, how do I remove the liquid from my plate?
Do NOT invert the plate to dump the liquid. The beads will not adhere to the plate. After centrifugation using a swing bucket rotor with a plate adaptor, you can remove the liquid by using a pipette.
- Is special software required for data analysis?
Typically flow cytometers generate output files in FCS format (e.g. FCS 2.0, 3.0, or 3.1) and in some cases in list mode file format (LMD). FCS files can be analyzed by different software. Data generated from using LEGENDplex™ kits can be analyzed using the LEGENDplex™ data analysis software freely available on the BioLegend website (biolegend.com/legendplex/software). A free of charge software license dongle is currently provided in the kit. Please check our website for the most updated versions of the software.
- Should I perform the assay with the filter plates or with V-bottom plates?
Filter plates or V-bottom plates have been included in some kits for your convenience. Performing the assay with filter plates is generally recommended. A vacuum filtration unit is required to work with the filter plates. If however, you don’t have access to a vacuum manifold, then you can use the V-bottom plates or micro-FACS tubes as described in the manual and follow the recommended assay protocols for the type of plates or tubes you choose. All tubes and plates should be made from low binding polypropylene. Polystyrene ELISA or cell culture plates should not be used.
- What is the shelf life of LEGENDplex™ kits?
LEGENDplex™ kits are valid for 6 months from the date of receipt.