Human Fas Ligand/TNFSF6 ELISpot Development Module, 5 Plate
Human Fas Ligand/TNFSF6 ELISpot Development Module, 5 Plate Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
PRODUCT SUMMARY
Complete ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. Kits are available for detection and enumeration of a single analyte or two analytes simultaneously. Complete ELISpot kits are ready-to-run and require no assay development or refinement. ELISpot Development Modules contain the basic components required to develop an ELISpot assay. They offer an economical alternative to buying separate antibodies.
ELISpot development modules are an alternative to ELISpot kits. A basic understanding of ELISpot assay development is required for the successful use of these reagents. Each investigator should optimize the coating conditions, the assay sensitivity, the type of enzyme and substrate, as well as the concentrations of the capture and detection antibodies to achieve desired results. The analyte-specific ELISpot Development Module and the ELISpot Blue Color Module contain the necessary components for analyte detection and visualization, respectively. These modules can be used together but are sold separately. Each module contains enough reagents for at least five 96-well microplates.
PRODUCT FEATURES
- An economical alternative to ELISpot Kits
- Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
- Generic development protocols provide direction to start an optimization protocol
- Customize the assay to your specific needs
MODULE CONTENTS
- Human Fas Ligand/TNFSF6 Capture Antibody
- Human Fas Ligand/TNFSF6 Biotinylated-conjugated Detection Antibody
OTHER REAGENTS REQUIRED
- ELISpot Blue Color Module or equivalent (R&D Systems, Catalog # SEL002)
- PBS - 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
- Wash Buffer - 0.05% Tween® 20 in PBS
- Blocking Buffer - 1% BSA, 5% Sucrose in PBS
- Reagent Diluent - 1% BSA in PBS, pH 7.2 - 7.4, 0.2 µm filtered
- 2 °C – 8 °C refrigerator
- 37 °C CO2 incubator
- Positive Control - Use Recombinant Human Fas Ligand/TNFSF6 or cells known to secrete Human Fas Ligand/TNFSF6
- 96-well plates - Nitrocellulose-bottom plates, PVDF-bottom Immunospot® plates, or flat-bottom polystyrene Immulon® ELISA plates
- Multi-channel pipette, squirt bottle, manifold dispenser, or automated microplate washer
- Dissection microscope or an automated ELISpot Reader
- Deionized H2O
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Product Datasheets
Preparation and Storage
Background: Fas Ligand/TNFSF6
Fas Ligand (FasL), also known as CD178, CD95L, or TNFSF6, is a transmembrane protein homotrimer that binds to Fas/CD95 and triggers apoptosis in the Fas-expressing cell. Fas Ligand also binds the soluble decoy receptor DcR3. It is expressed on activated Th1 cells, CD8+ cytotoxic T cells, and NK cells. A soluble form can be released which remains trimeric and retains the ability to bind Fas. Fas Ligand-induced apoptosis plays a central role in the development of immune tolerance and the maintance of immune privileged sites. Tumor cells can evade immune surveillance by upregulating Fas Ligand to kill tumor infiltrating lymphocytes. In gld mice, a Fas Ligand point mutation is the cause of severe lymphoproliferation and systemic autoimmunity.
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