Second-Messenger Cyclic AMP Plays an Unusual Role in T Cell Receptor Activation
| Cyclic adenosine monophosphate (cAMP) is an important second-messenger in many signal transduction pathways linking activation of cell surface membrane receptors to intracellular responses and, ultimately, to changes in gene expression. The second-messenger cAMP plays a rather complicated role in T cell receptor activation. T cell receptor activation stimulates cAMP production; however, cAMP as part of a negative feedback loop inhibits T cell function and proliferation (Mol Cell Biol 30:1660–1672 (2010)).
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Molecular Probes® Calcium Indicators for Studying T Cell Activation
| Ion-sensitive probes have enabled researchers to investigate changes in intracellular free Ca2+ concentrations using fluorescence microscopy, flow cytometry, and fluorescence spectroscopy. The properties and applications of these fluorescent indicators—most of which are derivatives of the Ca2+ chelators EGTA, APTRA, and BAPTA—have been extensively reviewed in the literature.
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| Product | Quantity | Cat. No. |
|---|---|---|
| X-rhod-1, AM, cell permeant, special packaging | 10 x 50 µg | X-14210 |
| Calcium Green™-1, AM, cell permeant, special packaging | 10 x 50 µg | C-3012 |
| Calcium Orange™, tetrapotassium salt, cell impermeant | 500 μg | C-3013 |
| Calcium Crimson™, AM, cell permeant, special packaging | 10 x 50 µg | C-3018 |
| Calcium Yellow™, tetrapotassium salt, cell impermeant | 500 µL | C36202 |
| Fluo-4, AM, cell permeant, special packaging | 10 x 50 µg | F-14201 |
Fluorogenic Substrate for the Detection of Activated Caspases
| The CellEvent™ Caspase-3/7 Green Detection Reagent comprises a DEVD peptide sequence conjugated to a nucleic acid–binding dye. In the presence of activated caspases-3 and -7, the CellEvent™ reagent fluoresces bright green (monitored with a standard FITC filter set, see figure). This robust assay enables researchers to examine caspase-3 and -7 activation in live cells. Furthermore, since wash steps are not necessary for detection, fragile apoptotic cells typically lost during wash steps are retained. Importantly, the fluorescent signal from CellEvent™ Caspase-3/7 Detection Reagent can survive fixation and permeabilization. This allows flexibility to perform endpoint assays and to probe the sample for other proteins of interest using immunocytochemistry.
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| Product | Qty. | Cat. No. |
|---|---|---|
| CellROX™ Deep Red Reagent, for oxidative stress detection | 5 x 50 µL | C10422 |
| CellEvent™ Caspase-3/7 Green Detection Reagent, 2 mM solution in DMSO | 100 µL | C10423 |
| Image-iT® DEAD Green Viability Stain | 25 µL | I10291 |
| Image-iT® LIVE Green Reactive Oxygen Species Detection Kit | 1 kit | I36007 |
| MitoSOX™ Red Mitochondrial Superoxide Indicator, for live-cell imaging | 5 x 50 µg | M36008 |
| Hoechst 33342 nucleic acid stain | 10 mL | H3570 |
How Does FOXP3 Regulate T Cells in Adaptive Immunity?
| Forkhead box-O transcription factors control expression of immune system genes in regulatory T cells. FOXP3 induces immunosuppressive functions in regulatory T cells and is a potential therapeutic target in tumor cells. ABfinity™ recombinant monoclonal antibodies are ideal tools for signaling pathway investigations. The ABfinity™ technology helps ensure consistent antibody performance, so you don’t have to reoptimize your assay with each new lot.
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| Product | Application* | Species | Qty. | Cat. No. |
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| FOXP3 ABfinity™ Recombinant Rabbit Monoclonal Antibody - Purified | ICC | Human | 100 µg | 700020 |
| FOXP3 ABfinity™ Recombinant Rabbit Monoclonal Antibody - Purified | WB | Human | 100 µg | 700914 |
| *ICC=Immunocytochemistry; WB=Western Blot | ||||
Alexa Fluor® Secondary Antibodies Light Up Your Pathway
| Stimulation of the T cell receptor pathway is triggered by major histocompatibility complex (MHC) molecules on antigen-presenting cells; this stimulation induces a series of intracellular signaling cascades that ultimately result in cellular proliferation, differentiation, cytokine production, and/or activation-induced cell death. |
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| Product | Qty. | Cat. No. |
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| Alexa Fluor® 488 goat anti-rabbit IgG (H+L), 2 mg/mL | 0.5 mL | A11008 |
| ZAP-70 [pY315/pY319] ABfinity™ Recombinant Rabbit Monoclonal Antibody | 100 µg | 700177 |
Measuring IL-2 Secretion After T Cell Activation
| T cell receptors are found on the surface of T lymphocytes. When an antigen or mitogen binds to these receptors and activates the mature T lymphocyte, transcription and release of synthesized IL-2 occurs.
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| Product | Qty. | Cat. No. |
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| Human IL-2 ELISA Kit | 96 tests | KHC0021 |
| Mouse IL-2 ELISA Kit | 96 tests | KMC0021 |
| Rat IL-2 ELISA Kit | 96 tests | KRC0021 |
| Monkey IL-2 ELISA Kit | 96 tests | KPC0021 |
T Cell Cytokine Signaling—Multiplex Luminex® Assays
| The functional aspects of many T cells are tied to the release of cytokines. Cytokines play an important role in T cell activation and are potent T cell growth factors, and many cytokines are secreted by helper T cells. T cell subsets have also been found to produce cytokines differentially in response to diverse stimuli.
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