Direct ELISA analysis of TNF alpha was performed by coating wells of a 96-well plate with 100ul per well of human recombinant protein TNF alpha (Product #RTNFAI) diluted in carbonate/bicarbonate buffer (Product # 28382) at a concentration of 1ug/ml overnight at 4C. Wells of the plate were washed, blocked with starting blocking buffer (Product # 37538), and incubated with 100ul per well of a mouse anti-TNF alpha monoclonal antibody (Product # MTNFAI) at a dilution of 1:25, 1:50, 1:100, 1:200, 1:400, 1:800, 1:1600 and 1:3200 for 90 minutes at 37C. The plate was washed, then incubated with 100ul per well of a HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 62-6520) at a dilution of 1:2000 for 90 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5-10 minutes at room temperature in the dark. The reaction was stopped with Stop solution (Product #N600), and absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human TNF alpha from Pichia pastoris.|
|Storage buffer||PBS with 5% trehalose|
|Tested Applications||Dilution *|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|ELISA (ELISA)||See 1 publications below|
MTNFAI targets TNF Alpha in ELISA, and Western blot applications and shows reactivity with Human samples. The MTNFAI antibody (clone E10043) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M3TNFABI (biotinylated conjugate of clone E10041).
The MTNFAI immunogen is recombinant human TNF alpha from Pichia pastoris.
MTNFAI detects TNF Alpha which has a predicted molecular weight of approximately 26 kDa.
This gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested the neuroprotective function of this cytokine.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Time-Resolved Fluorometric Method for One-Step Immunoassays Using Plasmonic Nanostructures.
MTNFAI was used in ELISA to evaluate a novel immunoassay with plasmonic nanostructures
|Szmacinski H,Murtaza Z,Lakowicz JR||The journal of physical chemistry. C, Nanomaterials and interfaces (114:7236)||2010|