- Other Names
- Multiplex assay, Flow cytometry, Inflammation, Flex set, Cytokines
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- Product Citations
Cytokines, which are soluble extracelluar proteins or glycoproteins, are key intercellular regulators and mobilizers of cells. They are secreted by immune cells and act on other cells to coordinate appropriate immune responses. They are crucial to innate and adaptive host defenses, cell growth, cell death, differentiation, as well as development and repair processes.
The Human Cytokine Panel 2 is a multiplex bead-based assay panel, using fluorescence–encoded beads suitable for use on various flow cytometers. This panel allows simultaneous quantification of 13 human cytokines, including TSLP, IL-1α, IL-1β, GM-CSF, IFN-α2, IL-23, IL-12p40, IL-12p70, IL-15, IL-18, IL-11, IL-27, and IL-33. This assay panel provides higher detection sensitivities and broader dynamic ranges than traditional ELISA methods. The panel has been validated by detecting expected changes in biological samples.
A LEGENDplex™ Mix and Match system is available to customize assays for specificities within this cytokine 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
- Detection Antibody (3.5 ml)
- Standard Cocktail, Lyophilized (1 vial)
- SA-PE (3.5 ml)
- Matrix B, Lyophilized (1 vial)
- Assay Buffer (25 ml)
- Wash Buffer (25 ml)
- Filter Plate (1 plate)
- Plate Sealers (4 sheets)
- Data Analysis Software Dongle
Learn more about LEGENDplex™ at biolegend.com/legendplex.
Download the LEGENDplex™ software here.
- Application Notes
Note: The target analyte IFN-α2 was previously listed as IFN-α. We have since determined that the antibody reagents used in this product are specific to only IFN-α2, and not all IFN-α isoforms.
- Materials Not Included
- Flow Cytometer
- Pipettes and Tips
- Reagent Reservoirs for Multichannel Pipettes
- Polypropylene Microfuge Tubes
- Vortex Mixer
- Plate Shaker
- A Vacuum Filtration Unit and a Vacuum Source (if using filter plates)
- Biology Area
- Cell Biology, Neuroinflammation, Neuroscience, Innate Immunity, Immunology
- Molecular Family
- Gene ID
- 3553 View all products for this Gene ID 3552 View all products for this Gene ID 3593 View all products for this Gene ID 3592 View all products for this Gene ID 1437 View all products for this Gene ID 3593 View all products for this Gene ID 3600 View all products for this Gene ID 3606 View all products for this Gene ID 85480 View all products for this Gene ID 3589 View all products for this Gene ID 246778 View all products for this Gene ID 90865 View all products for this Gene ID
- 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.