- Regulatory Status
- Other Names
- IL-10, IFN-γ, TNF-α, IL-4, IL-6, IL10A, Cachectin, Interferon Beta-2
- Ave. Rating
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- Product Citations
Upon antigen presentation by APCs (Antigen Presenting Cells), naive CD4+ T cells can differentiate into one of several lineages of T helper (Th) cells, including Th1, Th2, Th17, Th9, Tfh, Th22, and ThGM as defined by their patterns of cytokine production and functions. T helper cells play a critical role in mediating adaptive immunity to a variety of pathogens. They secrete cytokines to stimulate various effector cells, such as cytotoxic T cells, B cells and macrophages. Accurate measurement of Th cytokine expression is critical in understanding the nature of the immune response and many disease processes.
The LEGENDplex™ Rat Tfh 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 5 rat cytokines, including IL-10, IFN-γ, TNF-α, IL-4, and IL-6, which are collectively secreted by Th1, Th2, Th9, Th17, Th22, ThGM, and Tfh cells. This assay panel provides higher detection sensitivity and broader dynamic range than traditional ELISA method. The panel has been validated for use on cell culture supernatant, serum and plasma samples. 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)
- Rat Tfh Panel V02 (5-plex) Capture Beads
- Rat Th Cytokine Panel Detection Abs V02 (3.5 mL)
- Rat Th Cytokine Panel Standard A Cocktail, Lyophilized (1 vial)
- Rat Th Cytokine Panel Standard B Cocktail, Lyophilized (1 vial)
- LEGENDplex™ SA-PE (3.5 mL)
- LEGENDplex™ Matrix C, Lyophilized (1 vial)
- LEGENDplex™ Lyophilized Standard Reconstitution Buffer (5 mL)
- LEGENDplex™ Assay Buffer (25 mL)
- LEGENDplex™ Wash Buffer (25 mL)
- Plate Sealers (4 sheets)
- V-bottom Plate
- Verified Reactivity
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)
- Centrifuge with a swinging bucket adaptor for microtiter plates (if using V-bottom plates)
- 1.1 mL polypropylene micro FACS tubes, in 96-tube rack
- Biology Area
- Adaptive Immunity, Cell Biology, Immunology
- Molecular Family
- Gene ID
- 3586 View all products for this Gene ID 25712 View all products for this Gene ID 24835 View all products for this Gene ID 287287 View all products for this Gene ID 24498 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.