Rat TNF-alpha DuoSet ELISA
R&D Systems, part of Bio-Techne | Catalog # DY510
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
62.5-4000 pg/mL
Sample Type
Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Rat
Rat TNF-alpha DuoSet ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Product Summary for Rat TNF-alpha DuoSet ELISA
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant rat TNF-alpha. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
100 µL
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Rat TNF-alpha DuoSet ELISA
Rat TNF-alpha ELISA Standard Curve
Kit Contents for Rat TNF-alpha DuoSet ELISA
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
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 DuoSet ELISA
Product Specific Notices for Rat TNF-alpha DuoSet ELISA
For research use only
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