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.
Key Product Details
Assay Length
Sample Type & Volume Required Per Well
Sensitivity
Assay Range
Product Summary for Rhesus Macaque TNF-alpha Quantikine ELISA Kit
Product Specifications
Assay Type
Format
Measurement
Detection Method
Conjugate
Species
Specificity
Cross-reactivity
Interference
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, EDTA Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 61 | 123 | 492 | 56 | 111 | 465 |
| Standard Deviation | 3.6 | 7.4 | 35.7 | 5.6 | 7.2 | 41.8 |
| CV% | 5.9 | 6.0 | 7.3 | 10.0 | 6.5 | 9.0 |
Recovery for Rhesus Macaque TNF-alpha Quantikine ELISA Kit
The recovery of rhesus macaque TNF-alpha spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Supernates (n=4) | 110 | 104-120 |
| EDTA Plasma (n=3) | 94 | 89-99 |
| Serum (n=4) | 98 | 90-109 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with various concentrations of rhesus macaque TNF-alpha in each matrix were diluted with Calibrator \line Diluent and then assayed.
Scientific Data Images for Rhesus Macaque TNF-alpha Quantikine ELISA Kit
Rhesus Macaque TNF-alpha ELISA Standard Curve
Preparation and Storage
Shipping
Stability & Storage
Background: TNF-alpha
Long Name
Alternate Names
Entrez Gene IDs
Gene Symbol
Additional TNF-alpha Products
Product Documents for Rhesus Macaque TNF-alpha Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Rhesus Macaque TNF-alpha Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
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Protocols
View specific protocols for Rhesus Macaque TNF-alpha Quantikine ELISA Kit (RHMTA0):
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for Rhesus Macaque TNF-alpha Quantikine ELISA Kit
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Q: Assay Diluent RD1-63 contains visible precipitate. Is this acceptable to use?
A: A small amount of precipitate is expected and acceptable in Assay Diluent RD1-63. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.
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
