- Regulatory Status
- Other Names
- IL-12p70, TNF-α, IL-6, IL-4, IL-10, IL-1β, Arginase, TARC, IL-1RA, IL-12p40, IL-23, IFN-γ, IP-10
- Ave. Rating
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- Product Citations
Macrophages are derived from the differentiation of monocytes in response to an infection or tissue damage. Their primary function is to recognize, engulf, and destroy pathogens, dying or dead cells and cellular debris. Macrophages are also professional antigen presenting cells that play a crucial role in modulating the host response, secreting an array of cytokines which aid in host defense, tissue repair, and immunoregulation.
The LEGENDplex™ Human Macrophage/Microglia Panel (13-plex) is a bead-based multiplex assay panel, which uses fluorescence-encoded beads suitable for use on various flow cytometers. It allows for simultaneous quantification 13 key targets involved in monocyte differentiation and macrophage function including IL-12p70, TNF-α, IL-6, IL-4, IL-10, IL-1β, Arginase, TARC, IL-1RA, IL-12p40, IL-23, IFN-γ, and IP-10. 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, serum, and plasma samples.
The LEGENDplex™ Human Macrophage/Microglia Panel is designed to allow flexible customization within the panel. It can also be divided into sub-panels such as:
LEGENDplex™ Human M1 Macrophage Panel (8-plex)
LEGENDplex™ Human M2 Macrophage Panel (6-plex)
LEGENDplex™ Human M1/M2 Macrophage Panel (10-plex)
Please visit www.biolegend.com/legendplex for more information on how to mix and match within the panel.
- 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 (varies, see manual)
- Detection Antibodies (3.5 ml)
- Standard Cocktail, Lyophilized (1 vial)
- SA-PE (3.5 ml)
- Matrix B6, Lyophilized (1 vial)
- Assay Buffer (25 ml)
- Wash Buffer, 20X (25 ml)
- Plate Sealers (4 sheets)
- V-bottom Plate
Please note: Lots manufactured before September 2021 may include a different version of the “Matrix B.” There is no change in the protocol. Simply use the Matrix reagent you received as suggested in the current manual.
- Verified Reactivity
Learn more about LEGENDplex™ at biolegend.com/legendplex
Download the LEGENDplex™ software here.
- Additional Product Notes
- Product Citations
- Materials Not Included
- Flow Cytometer
- Pipettes and Tips
- Reagent Reservoirs for Multichannel Pipettes
- Polypropylene Microfuge Tubes
- Vortex Mixer
- Aluminum Foil
- Absorbent Pads or Paper Towels
- Plate Shaker
- Tabletop Centrifuges
- A Vacuum Filtration Unit and a Vacuum Source (if using filter plates)
- Biology Area
- Immunology, Innate Immunity, Neuroinflammation, Neuroscience
- Molecular Family
- Gene ID
- If I don't have a vacuum, how do I remove the liquid from my plate?
If you do not have a vacuum, the assay should be run in a V-bottom plate. After centrifugation using a swinging-bucket rotor with a plate adaptor, you can remove the liquid by flicking the plate quickly, dumping the contents into a sink, and patting it dry carefully on a stack of clean paper towels without losing the beads. Alternatively, you can remove the liquid by using a pipette.
- 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. A vacuum filtration unit is required to work with the filter plates. However, if you don’t have access to a vacuum manifold or if you prefer, then you can use the V-bottom plates and follow the recommended assay protocols for the type of plates you choose. All plates should be made from low binding polypropylene. Polystyrene ELISA or cell culture plates should not be used.
- 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.
- What is the shelf life of LEGENDplex™ kits?
LEGENDplex™ kits are guaranteed for 6 months from the date of receipt, but may have a shelf life of up to 2 years from the date of manufacture.
- 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). Other software may be available to analyze FCS files. Data generated using LEGENDplex™ kits can be analyzed using the freely available LEGENDplex™ data analysis software. Please check our website for the most updated versions of the software.