Rat TNF-alpha Quantikine ELISA Kit
R&D Systems, part of Bio-Techne | Catalog # RTA00
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (17 µL), Serum (25 µL), EDTA Plasma (25 µL), Heparin Plasma (25 µL)
Sensitivity
5 pg/mL
Assay Range
12.5-800 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Product Summary for Rat TNF-alpha Quantikine ELISA Kit
The Quantikine Rat TNF-alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure rat TNF-alpha levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant rat TNF-alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant rat TNF-alpha accurately. Results obtained using natural rat 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 rat TNF-alpha.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Rat
Specificity
Natural and recombinant rat 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.
Cell Culture Supernates, 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) | 65 | 232 | 593 | 63 | 246 | 656 |
| Standard Deviation | 3.3 | 5.1 | 12.5 | 6.1 | 23.6 | 57.6 |
| CV% | 5.1 | 2.2 | 2.1 | 9.7 | 9.6 | 8.8 |
Recovery for Rat TNF-alpha Quantikine ELISA Kit
The recovery of rat 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) | 101 | 93-119 |
| EDTA Plasma (n=5) | 96 | 88-104 |
| Heparin Plasma (n=4) | 80 | 73-84 |
| Serum (n=8) | 93 | 83-105 |
Linearity
To assess the linearity of the assay, samples spiked with various concentrations of rat TNF-alpha were diluted with Calibrator Diluent and then assayed. Results from typical sample dilutions are shown.
Scientific Data Images for Rat TNF-alpha Quantikine ELISA Kit
Rat 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
Rat 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 156 aa extracellular
domain (ECD) (12). Within the ECD, rat TNF-alpha shares 95% aa
sequence identity with mouse, and
73% - 79% aa identity with bovine, canine, cotton rat,
equine, feline, human, rhesus macaque,
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 of
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 NF kappaB 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-alpha (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 Rat TNF-alpha Quantikine ELISA Kit
Product Specific Notices for Rat TNF-alpha Quantikine ELISA Kit
For research use only
Loading...
Loading...
Loading...
⚠ 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.