Human TNF-alpha ELISpot Development Module, 5 Plate

R&D Systems | Catalog # SEL210

R&D Systems
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Key Product Details

Species

Human

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 (plus plate coating)

Sample Type

Whole Cells

Product Summary for Human TNF-alpha ELISpot Development Module, 5 Plate


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

Trademarks and registered trademarks are the property of their respective companies.

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Formulation, Preparation, and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

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

TNF-alpha (Tumor necrosis factor alpha) plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism. TNF-alpha was first identified as a cytotoxic factor produced by macrophages capable of killing mouse tumor cells. It is the prototypic ligand and along with Lymphotoxin-alpha, were identified as the first members of the TNF superfamily. Active TNF-alpha and other members of the TNF superfamily exist as a homotrimer with high structural homology. Receptor binding occurs at the interface of two TNF-alpha monomers. And receptor activation occurs when all three monomer interfaces are engaged with a receptor. For TNF-alpha, receptor binding and activation occurs through TNF R1 or TNF RII, and subsequently leads to activation of NF-kB or MAPK signaling pathways. Another pathway that TNF-alpha can activate utilizes the death domain of TNF RI to induce apoptosis. TNF-alpha promotes the inflammatory response largely through NF-kB signaling, and inhibition of TNF-alpha has proven successful in treating many autoimmune disorders. TNF-alpha is also present on the cell surface as membrane-bound TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells. TNF-alpha protein is translated as a type II transmembrane protein containing an N-terminal transmembrane domain. The soluble cytokine is released from its cell-anchoring TM domain by proteolytic processing by metalloproteases.

Long Name

Tumor Necrosis Factor alpha

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Gene Symbol

TNF

Additional TNF-alpha Products

Product Documents for Human TNF-alpha ELISpot Development Module, 5 Plate

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human TNF-alpha ELISpot Development Module, 5 Plate

For research use only

Citations for Human TNF-alpha ELISpot Development Module, 5 Plate

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