Porcine TNF-alpha ELISpot Development Module, 5 Plate

Catalog #: SEL690 Datasheet / COA / SDS

Discontinued Product

SEL690 has been discontinued.
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R&D Systems ELISpot Development Modules
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Porcine TNF-alpha ELISpot Development Module, 5 Plate Summary

Assay Type
Quantitative Sandwich ELISA Development Module
Format
96-well microplate, sold separately (see Other Reagents Required)
Assay Length
3 hours 35 mins to 4 hours 50 mins**
Sample Type
Whole Cells
Sufficient Materials
For five 96-well microplates
Specificity
Please see the product datasheet

* 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

  • Porcine TNF-alpha Capture Antibody
  • Porcine TNF-alpha 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 Porcine TNF-alpha or cells known to secrete Porcine TNF-alpha
  • 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

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: TNF-alpha

Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71% - 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness.

Cleavage of membrane bound TNF-α by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.

Long Name:
Tumor Necrosis Factor alpha
Entrez Gene IDs:
7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
Alternate Names:
APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha

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