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.
Mouse TNF-alpha ELISpot Kit
R&D Systems | Catalog # EL410
Key Product Details
Species
Assay Type
Format
Assay Length
Sample Type
Product Summary for Mouse TNF-alpha ELISpot Kit
PRODUCT SUMMARY
ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. This kit is designed for the detection and enumeration of mouse TNF-alpha. Complete ELISpot kits are ready-to-run and require no assay development or refinement.
This ELISpot assay employs a capture antibody specific for mouse TNF-alpha, pre-coated onto a PVDF-backed microplate. Appropriately stimulated cells are pipetted directly into the wells and the immobilized antibody in the immediate vicinity of the secreting cells binds secreted mouse TNF-alpha. Following wash steps and incubation with a biotinylated detection antibody, alkaline-phosphatase conjugated to streptavidin is added. Unbound enzyme is subsequently removed by washing and a substrate solution (BCIP/NBT) is added. A blue-black colored precipitate forms at the sites of cytokine localization and appears as spots, with each individual spot representing an individual mouse TNF-alpha secreting cell. The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.
PRODUCT FEATURES
- Detect and quantitate individual cells secreting mouse TNF-alpha
- High sensitivity - ELISpot assays can measure responses with frequencies well below 1 in 100,000 cells
- No in vitro expansion of cells required
- High-throughput - ELISpot assays use only a small number of primary cells
KIT CONTENTS
- Mouse TNF-alpha Microplate
- Biotinylated Detection Antibody
- Streptavidin conjugated to Alkaline Phosphatase
- Dilution Buffers
- Wash Buffer Concentrate
- BCIP/NBT Chromogen
- Mouse TNF-alpha Positive Control
OTHER REAGENTS REQUIRED
- Pipettes and pipette tips
- Deionized or distilled water
- Squirt bottle, manifold dispenser, or automated microplate washer
- 500 mL graduated cylinder
- 37 °C CO2 incubator
- Sterile culture media
- Dissection microscope or an automated ELISpot reader
Formulation, Preparation, and Storage
Shipping
Storage
Background: TNF-alpha
Long Name
Alternate Names
Entrez Gene IDs
Gene Symbol
Additional TNF-alpha Products
Product Documents for Mouse TNF-alpha ELISpot Kit
Product Specific Notices for Mouse TNF-alpha ELISpot Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including methanol, which is known to the State of California to cause reproductive toxicity with developmental effects. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Mouse TNF-alpha ELISpot Kit
Powered by Bioz
Customer Reviews for Mouse TNF-alpha ELISpot Kit
There are currently no reviews for this product. Be the first to review Mouse TNF-alpha ELISpot Kit and earn rewards!
Have you used Mouse TNF-alpha ELISpot Kit?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Associated Pathways
Apoptosis Signaling Pathway
Dendritic Cell Lineage Development Pathways
Hematopoietic Stem Cell Differentiation Pathways & Lineage-specific Markers
IL-2 Signaling Pathways and their Primary Biological Effects in Different Immune Cell Types
Innate Lymphoid Cell Differentiation Pathways
mTOR Signaling Pathway
NOD-like Receptor Signaling Pathways
Th1 Differentiation Pathway
TNF Superfamily Pathway: Human Ligand-Receptor Interactions & their Associated Functions
Toll-Like Receptor Signaling Pathways
