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.
Human TNF-alpha Quantikine HS ELISA
R&D Systems | Catalog # HSTA00D
Key Product Details
Assay Length
6.5 hours
Sample Type & Volume Required Per Well
Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL), Citrate Plasma (200 µL)
Sensitivity
0.191 pg/mL
Assay Range
0.5-32 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Loading...
Product Summary for Human TNF-alpha Quantikine HS ELISA
The Quantikine HS Human TNF-alpha Immunoassay is a 6.5 hour solid phase ELISA designed to measure TNF-alpha in serum and plasma. It contains E. coli-derived recombinant human TNF-alpha and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human TNF-alpha. Results obtained with naturally occurring TNF-alpha samples showed linear curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural TNF-alpha. Since the measurement of TNF-alpha is insensitive to the addition of recombinant forms of either of the two types of soluble receptors, it is probable that this measurement detects the total amount of TNF-alpha in samples, i.e., the total amount of free TNF-alpha plus the amount of TNF-alpha bound to soluble receptors.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450 or 490nm (TMB or AP Substrate)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human TNF-alpha
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
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. Assays were performed by at least three technicians using two lots of components.
Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 41 | 41 | 41 |
| Mean (pg/mL) | 1.96 | 11.5 | 22.1 | 1.83 | 10.5 | 20.3 |
| Standard Deviation | 0.17 | 0.49 | 0.68 | 0.19 | 0.76 | 1.50 |
| CV% | 8.7 | 4.3 | 3.1 | 10.4 | 7.2 | 7.4 |
Recovery for Human TNF-alpha Quantikine HS ELISA
The recovery of TNF-alpha was determined by spiking to three different levels throughout the range of the assay in various matrices.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Citrate Plasma (n=4) | 95 | 88-105 |
| EDTA Plasma (n=4) | 99 | 92-111 |
| Heparin Plasma (n=4) | 93 | 87-103 |
| Serum (n=4) | 93 | 85-98 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of TNF-alpha and diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human TNF-alpha Quantikine HS ELISA
Human TNF-alpha High Sensitivity Standard Curve
Human TNF-alpha Standard Curve
Human TNF-alpha Quantikine HS ELISA Kit Sensitivity Competitor Comparison
Human TNF-alpha Quantikine HS ELISA Kit Recovery Competitor Comparison
Human TNF-alpha ELISA Standard Curve
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 2 - 8 °C. Do not use past expiration date.
Background: TNF-alpha
Long Name
Tumor Necrosis Factor alpha
Alternate Names
Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2
Entrez Gene IDs
Gene Symbol
TNF
Additional TNF-alpha Products
Product Documents for Human TNF-alpha Quantikine HS ELISA
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 Human TNF-alpha Quantikine HS ELISA
For research use only
Related Research Areas
Citations for Human TNF-alpha Quantikine HS ELISA
Powered by Bioz
Customer Reviews for Human TNF-alpha Quantikine HS ELISA (3)
5 out of 5
3 Customer Ratings
Have you used Human TNF-alpha Quantikine HS ELISA?
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
Customer Images
Showing
1
-
3 of
3 reviews
Showing All
Filter By:
-
Sample Tested: EDTA PlasmaVerified Customer | Posted 01/18/2018
-
Sample Tested: Peripheral blood neutrophilsVerified Customer | Posted 08/04/2017Due to the short experimental time, the standard TNFa duoSET ELISA did not work, however HS kit did.
-
Sample Tested: Human serumVerified Customer | Posted 05/25/2016
There are no reviews that match your criteria.
Protocols
View specific protocols for Human TNF-alpha Quantikine HS ELISA (HSTA00D):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- 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 200 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
- Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 6 times.
- Add 50 µL Substrate Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour. Do not wash the plate.
- Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
- Add 50 µL of Stop Solution to each well. Read at 490 nm within 30 minutes. Set wavelength correction to 650 nm or 690 nm.
50 µL Assay Diluent
200 µL Standard, Control, or Sample
200 µL Conjugate
50 µL Substrate Solution
50 µL Amplifier Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
Loading...
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
IL-15 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
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