Figure 1. PBMCs were initially stimulated with immobilized anti-CD3 (20 mg/well), soluble anti-CD28 antibody (1 μg/ml), recombinant hIL-4 #8919 (1 ng/ml), and recombinant hIL-2 #8907 (2 ng/ml) for a total of 5 d. The cells were then washed and either untreated (-) or treated (+) with TPA #4174 (10 ng/ml) and ionomycin (1 μg/ml). Cell culture media supernatants were removed after 4 and 24 hr and hIL-2 levels were quantified using SignalKine™ Human IL-2 Chemiluminescent Sandwich ELISA Kit.Learn more about how we get our images
Typical Data. Represented above are hIL-2 Standard Curves, one diluted in SignalKine™ Sample Diluent S01, a second diluted in SignalKine™ Sample Diluent S03, and a third in cell culture media (RPMI + 10% FBS). Although these are typical of the standard curves that will be generated using this kit, a new set of standards should be run for each new experiment.Learn more about how we get our images
Sample TestingLearn more about how we get our images
|Product Includes||Volume||Solution Color|
|hIL-2 Mouse mAb Coated Microwells (Chemiluminescent)||96 tests|
|Human IL-2 Standard (Lyophilized)||1 vial|
|SignalKine™ Assay Diluent A01||1.5 ml||Colorless|
|SignalKine™ Sample Diluent S01||25 ml||Brown|
|SignalKine™ Sample Diluent S03||25 ml||Brown|
|hIL-2 Detection Rabbit mAb (Biotinylated)(Lyophilized) 100X||1 vial|
|Detection Antibody Diluent||6 ml||Green|
|HRP-linked Streptavidin (Lyophilized) 100X||1 vial|
|HRP Diluent||6 ml||Red|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|Luminol/Enhancer Solution||3 ml||Colorless|
|Stable Peroxide Buffer||3 ml||Colorless|
|Sealing Tape||2 ea|
SignalKine™ Human IL-2 Chemiluminescent Sandwich ELISA Kit from Cell Signaling Technology (CST) is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects human IL-2 (hIL-2) in multiple matrices. Unknown samples being tested for hIL-2 and hIL-2 standards are added to low volume microwells, where the hIL-2 is captured by the coated hIL-2 Mouse mAb. Following a washing step, a biotinylated hIL-2 Detection Rabbit mAb is added to detect the captured hIL-2. HRP-linked Streptavidin is then used for detection of the biotinylated hIL-2 Detection Rabbit mAb. A chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU) is proportional to the quantity of human hIL-2 in the sample. SignalKine™ Human IL-2 Chemiluminescent Sandwich ELISA Kit detects hIL-2 in multiple matrices that can be quantified by generating a standard curve with the recombinant hIL-2 protein standard provided. The hIL-2 standard range is from 31.2 to 2000 pg/ml. Samples containing higher levels of hIL-2 can be diluted to fit into the standards range.
Interleukin-2 (IL-2) is a T cell stimulatory cytokine best known for inducing T cell proliferation and NK cell proliferation and activation (1,2). IL-2 also promotes peripheral development of regulatory T cells (Tregs) (3,4). Conversely, IL-2 is involved in the activation-induced cell death (AICD) that is observed post T cell expansion by increasing levels of Fas on CD4+ T cells (5). The effects of IL-2 are mediated through a trimeric receptor complex consisting of IL-2Rα, IL-2Rβ, and the common gamma chain, γc (1,2). IL-2Rα binds exclusively to IL-2 with low affinity and increases the binding affinity of the whole receptor complex including IL-2Rβ and γc subunits. IL-15 also binds to IL-2Rβ (1,2). γc is used by other cytokines including IL-4, IL-7, IL-9, IL-15, and IL-21 (1,2). Binding of IL-2 initiates signaling cascades involving Jak1, Jak3, Stat5, and the PI3K/Akt pathways (1,2).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Explore pathways related to this product.