Mouse TNF-alpha Quantikine HS ELISA Kit
R&D Systems, part of Bio-Techne | Catalog # MHSTA50
Key Product Details
Assay Length
4.0 hours
Sample Type & Volume Required Per Well
Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)
Sensitivity
0.295 pg/mL
Assay Range
0.781-50 pg/mL (Serum, EDTA Plasma, Heparin Plasma)
Product Summary for Mouse TNF-alpha Quantikine HS ELISA Kit
The Quantikine HS Mouse TNF-alpha Immunoassay is a 4.0 hour solid phase ELISA designed to measure mouse TNF-alpha levels in serum and plasma. It contains E. coli-expressed recombinant mouse TNF-alpha and antibodies raised against the recombinant factor and has been shown to accurately quantitate recombinant mouse TNF-alpha. Results obtained using natural TNF-alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine HS kit standards. These results indicate that the Quantikine HS Immunoassay kit can be used to determine relative mass values for natural mouse TNF-alpha.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450 or 490nm (TMB or AP Substrate)
Conjugate
HRP
Reactivity
Mouse
Specificity
Natural and recombinant mouse 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 twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
EDTA Plasma, Heparin 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) | 6.63 | 15.1 | 29.2 | 6.29 | 15.2 | 28.5 |
| Standard Deviation | 0.257 | 0.695 | 1.49 | 0.552 | 1.18 | 1.94 |
| CV% | 3.9 | 4.6 | 5.1 | 8.8 | 7.8 | 6.8 |
Recovery for Mouse TNF-alpha Quantikine HS ELISA Kit
The recovery of mouse TNF-alpha spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| EDTA Plasma (n=4) | 91 | 81-104 |
| Heparin Plasma (n=4) | 90 | 79-106 |
| Serum (n=4) | 86 | 77-99 |
Linearity
To assess the linearity of the assay, samples spiked with high concentrations of mouse TNF-alpha were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Mouse TNF-alpha Quantikine HS ELISA Kit
Mouse TNF-alpha ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped with polar packs. 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
Mouse TNF-alpha is synthesized as a 26 kDa type II transmembrane
protein that consists of a
35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane
segment, and a 179 aa
extracellular domain (ECD) (12). Within the ECD, mouse TNF-alpha
shares 95% aa identity with rat,
and 80% aa identity with canine, equine, feline, human,
rabbit, and porcine TNF-alpha. It is
produced by a wide variety of immune, epithelial,
endothelial, and tumor cells. TNF-alpha is
assembled intracellularly to form a noncovalently linked
homotrimer which is expressed on the
cell surface (13). Cell surface TNF-alpha can both induce the
lysis of tumor cells and virus infected
cells, and generate its own downstream cell signaling
following ligation by soluble TNF RI
(14, 15). Shedding of membrane bound TNF-alpha by TACE/ADAM17
releases the bioactive
cytokine, a 55 kDa soluble trimer containing the TNF-alpha
extracellular domain (16-18).
TNF-alpha binds the ubiquitous 55-60 kDa TNF RI (19, 20) and the
hematopoietic cell-restricted
78-80 kDa TNF RII (21, 22), both of which are also expressed
as homotrimers (1, 23). Both type I
and type II receptors bind TNF-alpha with comparable affinity
and can promote NFkB activation
(24-27). Only TNF RI, however, contains a cytoplasmic death
domain which triggers the
activation of apoptosis (3, 28). Soluble forms of both types
of receptors are released into
human serum and urine, and can neutralize the biological
activity of TNF (29-31).
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 Mouse TNF-alpha Quantikine HS ELISA Kit
Product Specific Notices for Mouse TNF-alpha Quantikine HS ELISA Kit
For research use only
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⚠ 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.