Product Details
Catalog Number: ELH-TNFalpha-001
Applications: E
Type: Kit
Species Reactivity: Human
Downloads: Datasheet (pdf)
Product Sizes
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1 Plate Kit$379.00Add to Cart

Sensitive, Specific and Cost Effective

TNF-alpha (tumor necrosis factor-a) is secreted by macrophages, monocytes, neutrophils, T-cells, NK-cells following their stimulation by bacterial lipopolysaccharides. Human TNF-alpha is a non-glycosylated protein of 17.5 kDa and a length of 157 amino acid. TNF-alpha shows a wide spectrum of biological activities. It causes cytolysis and cytostasis of many tumor cell ines in vitro. Within hours after injection TNF-alpha leads to the destruction of small blood vessels within malignant tumors. TNF-alpha also enhances phagocytosis and cytotoxicity in neutrophilic granulocytes and also modulates the expression of many other proteins. In general, TNF-alpha and TNF-beta display similar spectra of biological activities in vitro systems although TNF-beta is often less potent or displays apparent partial agonist activity.

Figure: TNF-alpha standard curve

Suggested Application:Quantitative Protein Detection; Establishing Normal Range; Validation of Antibody Array Results.

Kit Components:
·   Pre-Coated 96-Well Strip Microplate
·   Wash Buffer
·   Stop Solution
·   Assay Diluent(s)
·   Lyophilized Standard
·   Biotinylated Detection Antibody
·   Streptavidin-Conjugated HRP
·   TMB One-Step Substrate
Other Materials Required:
·     Distilled or deionized water
·     Precision pipettes to deliver 2 μl to 1 ml volumes
·     Adjustable 1-25 ml pipettes for reagent preparation
·     100 ml and 1 liter graduated cylinders
·     Tubes to prepare standard and sample dilutions
·     Absorbent paper
·     Microplate reader capable of measuring absorbance at 450nm
·     Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline:
1.Prepare all reagents, samples and standards as instructed in the manual.
2.Add 100 μl of standard or sample to each well. Incubate 2.5 h at RT or O/N at 4°C.
3.Add 100 μl of prepared biotin antibody to each well. Incubate 1 h at RT.
4.Add 100 μl of prepared Streptavidin solution to each well. Incubate 45 min at RT.
5.Add 100 μl of TMB One-Step Substrate Reagent to each well. Incubate 30 min at RT.
6.Add 50 μl of Stop Solution to each well. Read at 450 nm immediately.

 

Figure: Box plot of the serum IL-6 level in the RA patients and control according to the IL-6 (−174 G/C) promoter polymorphism type. http://dx.doi.org/10.1016/j.ejr.2012.11.002

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