- Other Names
- Multiplex assay, flow cytometry, inflammation, T helper cell, CBA, cytokines, Th1, Th2, Th9, Th17, Th1/Th2, Th22
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T helper (Th) cells play important roles in regulating immune responses. They secrete cytokines to stimulate various effector cells, such as cytotoxic T cells, B cells and macrophages. These cytokines are also involved in inflammatory and autoimmune diseases. Accurate measurement of Th cytokine expression is critical for in-depth understanding of the immune responses and many disease processes.
The LEGENDplex™ Human Th Cytokine Panel is a bead-based multiplex assay panel, using fluorescence–encoded beads suitable for use on various flow cytometers. This panel allows simultaneous quantification of 12 human cytokines, including IL-2, 4, 5, 6, 9, 10, 13, 17A, 17F, 22, IFN-γ and TNF-α, which are collectively secreted by Th1, Th2, Th9, Th17, and Th22. This assay panel provides higher sensitivities and broader dynamic range than traditional ELISA methods. The panel has been validated for use on serum and cell culture supernatant samples.
The LEGENDplex™ Human Th Panel is designed to allow flexible customization within the panel. It can also be divided into sub-panels, such as:
LEGENDplex™ HU Th9 Panel (3-Plex)
LEGENDplex™ HU Th22 Panel (4-Plex)
LEGENDplex™ HU Th1 Panel (5-Plex)
LEGENDplex™ HU Th2 Panel (6-Plex)
LEGENDplex™ HU Th17 Panel (7-Plex)
LEGENDplex™ HU Th1/Th2 Panel (8-Plex)
Please refer to the target selection table for panel-specific target information (Table 1). Please visit www.biolegend.com/legendplex for more information on panel design and 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 B, Lyophilized (1 vial)
- Assay Buffer (25 mL)
- Wash Buffer, 20X (25 mL)
- Plate Sealers (4 sheets)
- Data Analysis Software Dongle
- V-bottom Plate
- Human Th Panel Manual
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
- 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)
- 1.1 mL polypropylene micro FACS tubes, in 96-tube rack
- Biology Area
- Cell Biology, Immunology, Neuroinflammation, Neuroscience
- Molecular Family
- Gene ID
- 3558 View all products for this Gene ID 3565 View all products for this Gene ID 3567 View all products for this Gene ID 3569 View all products for this Gene ID 3578 View all products for this Gene ID 3586 View all products for this Gene ID 3596 View all products for this Gene ID 3605 View all products for this Gene ID 112744 View all products for this Gene ID 50616 View all products for this Gene ID 3458 View all products for this Gene ID 7124 View all products for this Gene ID
- View information about Th Cytokine 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.