Human TNF-alpha DuoSet ELISA Best Seller
R&D Systems, part of Bio-Techne | Catalog # DY210
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
15.6-1000 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
Human
Human 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 Human 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 human 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 Human TNF-alpha DuoSet ELISA
Human TNF-alpha ELISA Standard Curve
Kit Contents for Human 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
Human 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 177 aa extracellular domain (ECD) (12, 13). Within the ECD,
human TNF-alpha shares 97% aa sequence identity with rhesus monkey, and 71%-92% aa
identity with bovine, canine, cotton rat, equine, feline, mouse, porcine, and
rat 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 (14). 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
(15, 16). Shedding of membrane bound TNF-alpha by TACE/ ADAM17 releases the
bioactive cytokine, a 55 kDa soluble trimer of the TNF-alpha extracellular domain
(17-19).
TNF-alpha binds the ubiquitous 55-60 kDa TNF RI (20, 21) and the
hematopoietic cell-restricted 78-80 kDa TNF RII (22, 23), both of which are
also expressed as homotrimers (1, 24). Both type I and type II receptors bind
TNF-alpha with comparable affinity and can promote NF kappaB activation (25-28). Only
TNF RI, however, contains a cytoplasmic death domain which triggers the
activation of apoptosis (3, 29). Soluble forms of both types of receptors are
released into human serum and urine and can neutralize the biological activity
of TNF-alpha (30-32).
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 DuoSet ELISA
Product Specific Notices for Human TNF-alpha DuoSet ELISA
For research use only
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