- Regulatory Status
- 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 B1, Lyophilized (1 vial)
- Assay Buffer (25 ml)
- Wash Buffer (25 ml)
- Lyophilized Standard Reconstitution Buffer (25 ml)
- Filter Plate (1 plate)
- Plate Sealers (4 sheets)
Please note: Lots manufactured after September 2021 may include a different version of the “Matrix B”. There is no change in the protocol, simply use Matrix B1 or any other version of Matrix B reagent as mentioned in the current manual.
Learn more about LEGENDplex™ at biolegend.com/legendplex
Download the LEGENDplex™ software here.
- Application Notes
Note: The target analyte IFN-a2 was previously listed as IFN-a. We have since determined that the antibody reagents used in this product are specific to only IFN-a2, and not all IFN-a isoforms.
- Product Citations
- 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, Immunology, Innate Immunity, Neuroinflammation, Neuroscience
- Molecular Family
- Gene ID
- 85480 View all products for this Gene ID 3552 View all products for this Gene ID 3553 View all products for this Gene ID 1437 View all products for this Gene ID 3439 View all products for this Gene ID 3440 View all products for this Gene ID 3593 View all products for this Gene ID 51561 View all products for this Gene ID 3592 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 3589 View all products for this Gene ID 246778 View all products for this Gene ID 90865 View all products for this 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.