The LEGENDplex™ HU Immune Checkpoint Panel 1 Standard contains 12 recombinant human proteins: sCD25 (IL-2Ra), 4-1BB, sCD27, B7.2 (CD86), free active TGF-β1, CTLA-4, PD-L1, PD-L2, PD-1, Tim-3, LAG-3, Galectin-9. This standard cocktail is recommended for use with the following in Mix and Match assays:
HU Immune Checkpoint Panel 1
740868 LEGENDplex™ HU Immune Checkpoint Panel 1 Detection Antibodies
740870 LEGENDplex™ Human sCD25 (IL-Ra) Capture Bead A4, 13X
740872 LEGENDplex™ Human 4-1BB Capture Bead A6, 13X
740873 LEGENDplex™ Human sCD27 Capture Bead A7, 13X
740874 LEGENDplex™ Human B7.2 (CD86) Capture Bead A8, 13X
740875 LEGENDplex™ Human Free Active TGF-β1 Capture Bead A10, 13X
740876 LEGENDplex™ Human CTLA-4 Capture Bead B2, 13X
740877 LEGENDplex™ Human PD-L1 Capture Bead B3, 13X
740878 LEGENDplex™ Human PD-L2 Capture Bead B4, 13X
740879 LEGENDplex™ Human PD-1 Capture Bead B5, 13X
740880 LEGENDplex™ Human Tim-3 Capture Bead B6, 13X
740881 LEGENDplex™ Human LAG-3 Capture Bead B7, 13X
740882 LEGENDplex™ Human Galectin-9 Capture Bead B9, 13X
740981 LEGENDplex™ Buffer Set M
740377 LEGENDplex™ Filter Plate
740379 LEGENDplex™ V-bottom Plate
- Biology Area
- Cancer Biomarkers, Immunology
- Molecular Family
- Cytokines/Chemokines, Soluble Receptors
- 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.