Mouse TNF-alpha ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # MTA00B
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)
Sensitivity
7.21 pg/mL
Assay Range
10.9-700 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Product Summary for Mouse TNF-alpha Quantikine ELISA Kit
The Quantikine Mouse TNF-alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse TNF-alpha levels in cell culture supernates, serum, and plasma. It contains E.
coli-expressed recombinant mouse TNF-alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse TNF-alpha accurately. Results obtained using natural mouse TNF-alpha showed dose-response 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 mouse TNF-alpha.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
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
Interference observed with 1 or more 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.
Cell Culture Supernates
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 54.6 | 116 | 405 | 52.6 | 138 | 373 |
| Standard Deviation | 1.50 | 4.51 | 12.5 | 4.61 | 8.49 | 29.7 |
| CV% | 2.7 | 3.9 | 3.1 | 8.8 | 6.2 | 8.0 |
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) | 50.5 | 134 | 375 | 53.0 | 146 | 388 |
| Standard Deviation | 1.91 | 4.95 | 11.5 | 3.69 | 8.42 | 29.7 |
| CV% | 3.8 | 3.7 | 3.1 | 7.0 | 5.8 | 7.7 |
Recovery for Mouse TNF-alpha Quantikine ELISA Kit
The recovery of mouse 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=6) | 104 | 94-111 |
| EDTA Plasma (n=4) | 95 | 85-103 |
| Heparin Plasma (n=4) | 92 | 86-98 |
| Serum (n=4) | 94 | 88-106 |
Linearity
To assess the linearity of the assay, samples were spiked with various concentrations of mouse TNF-alpha in each matrix, diluted with the appropriate Calibrator Diluent and then assayed.
Scientific Data Images for Mouse TNF-alpha Quantikine ELISA Kit
Mouse TNF-alpha ELISA Cell Culture Supernate Standard Curve
Mouse TNF-alpha ELISA Serum/Plasma Standard Curve
Mouse TNF-alpha ELISA Kit - Quantikine
HFD-induced plasma saturated free fatty acid (FFA) and lactate trigger production of proinflammatory cytokines and VEGF in mouse chondrocytes.(a~d) Relative mRNA levels of IL-6 (a), TNF-alpha (b), IL-1 beta (c) and VEGF (d) in the mouse primary chondrocytes treated with full or fractionated (>3 KD or <3 KD) serum from the 8-week ND or HFD-feeding mice. (n = 3, *P < 0.05, **P < 0.01 and ***P < 0.005). (e~h) Relative mRNA levels of IL-6 (e), TNF-alpha (f), IL-1 beta (g) and VEGF (h) in the mouse primary chondrocytes treated with BSA (5%) or BSA-linked FFA (18:0, 200 μM) for 24 h. (n = 3, *P < 0.05 and **P < 0.01). (i~l) Relative mRNA expression of IL-6 (i), TNF-alpha (j), IL-1 beta (k) and VEGF (l) in the mouse primary chondrocytes treated with lactate (25 mM) or vehicle control for 24 h. (n = 3, *P < 0.05 and **P < 0.01). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26271607), licensed under a CC-BY license. Not internally tested by R&D Systems.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
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 ELISA Kit
Product Specific Notices for Mouse TNF-alpha Quantikine 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.