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.
· 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
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
- Product Reviews
- Related Products
Name Catalog # Activin A Sandwich ELISA Kit ELH-Activin-A-001 Eotaxin-1/CCL11 Sandwich ELISA Kit ELH-Eotaxin-001 Eotaxin-1/CCL11 Sandwich ELISA Kit ELH-Eotaxin-002 IFN-gamma Sandwich ELISA Kit ELH-IFNgamma-001 IFN-gamma Sandwich ELISA Kit ELH-IFNgamma-002 IFN-gamma Sandwich ELISA Kit ELH-IFNgamma-003 IGFBP2 Sandwich ELISA Kit ELH-IGFBP2-001 IGFBP2 Sandwich ELISA Kit ELH-IGFBP2-002 IGFBP2 Sandwich ELISA Kit ELH-IGFBP2-003 Il-12p40 Sandwich ELISA Kit ELH-IL12p40-001 Il-12p40 Sandwich ELISA Kit ELH-IL12p40-002 Il-12p40 Sandwich ELISA Kit ELH-IL12p40-003 IL-6 Sandwich ELISA Kit ELH-IL6-001 IL-6 Sandwich ELISA Kit ELH-IL6-002 IL-6 Sandwich ELISA Kit ELH-IL6-003 MMP2 Sandwich ELISA Kit ELH-MMP2-001 MMP9 Sandwich ELISA Kit ELH-MMP9-001 MMP9 Sandwich ELISA Kit ELH-MMP9-002 MMP9 Sandwich ELISA Kit ELH-MMP9-003 TNF-alpha Human Sandwich ELISA Kit ELH-TNFalpha-002 TNF-alpha Human Sandwich ELISA Kit ELH-TNFalpha-003 IL-1 beta Mouse Sandwich ELISA Kit ELM-IL1b-001 IL-1 beta Mouse Sandwich ELISA Kit ELM-IL1b-002 IL-1 beta Mouse Sandwich ELISA Kit ELM-IL1b-003 TNF-alpha Mouse Sandwich ELISA Kit ELM-TNFalpha-001 TNF-alpha Mouse Sandwich ELISA Kit ELM-TNFalpha-002 TNF-alpha Mouse Sandwich ELISA Kit ELM-TNFalpha-003