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Human TNF-alpha ELISA - Quantikine HS Best Seller

R&D Systems, part of Bio-Techne | Catalog # HSTA00E

R&D Systems, part of Bio-Techne
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HSTA00E
PHSTA00E
SSTA00E

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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

0.049 pg/mL

Assay Range

0.156-10 pg/mL (Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Human TNF-alpha Quantikine HS ELISA

The Quantikine® HS Human TNF-alpha Immunoassay is a 4.0 hour solid phase ELISA designed to measure human TNF-alpha in serum and plasma. It contains E. coli-derived recombinant human TNF-alpha and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human TNF-alpha. Results obtained with natural human TNF-alpha samples showed linear curves that were parallel to the standard curves obtained using the Quantikine® kit standards. These results indicate that this kit can be used to determine relative mass values for natural human TNF-alpha.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450 or 490nm (TMB or AP Substrate)

Conjugate

HRP

Reactivity

Human

Specificity

Natural and recombinant human 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) 1.07 3.30 6.71 1.09 3.26 6.51
Standard Deviation 0.024 0.066 0.129 0.073 0.220 0.406
CV% 2.2 2.0 1.9 6.7 6.7 6.2

Recovery for Human TNF-alpha Quantikine HS ELISA

The recovery of human 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) 97 80-111
Heparin Plasma (n=4) 96 80-111
Serum (n=4) 94 83-103

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of human TNF-alpha were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

Human TNF-alpha High Sensitivity ELISA Linearity
Human TNF-alpha  ELISA Linearity
Human TNF-alpha  ELISA Linearity

Scientific Data Images for Human TNF-alpha Quantikine HS ELISA

Human TNF-alpha  Quantikine HS ELISA Kit Sensitivity Competitor Comparison

Human TNF-alpha Quantikine HS ELISA Kit Sensitivity Competitor Comparison

Human TNF-alpha  Standard Curve

Human TNF-alpha Standard Curve

Human TNF-alpha High Sensitivity Standard Curve

Human TNF-alpha High Sensitivity 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

Tumor Necrosis Factor alpha (TNF-alpha), also known as cachectin and TNFSF1A, is the prototypic ligand of the TNF superfamily (1). It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism (2-5). TNF-alpha is also involved in a number of pathological conditions including asthma, Crohn's disease, rheumatoid arthritis, neuropathic pain, obesity, type 2 diabetes, septic shock, autoimmunity, and cancer (5-11). 
 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

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Gene Symbol

TNF

Additional TNF-alpha Products

Product Documents for Human TNF-alpha Quantikine HS ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human TNF-alpha Quantikine HS ELISA

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.