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 biotin conjugated mouse anti-TNF alpha monoclonal antibody (Product # M3TNFABI) 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 pre-diluted High Sensitivity Streptavidin-HRP antibody (Product # 21134) at a dilution of 1:50 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||Dog, Cat, Human, Non-human primate, Pig|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human TNF alpha from Pichia pastoris.|
|Storage buffer||PBS with 5% trehalose|
|Storage Conditions||4° C|
|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|
M3TNFABI targets TNF Alpha in ELISA, and Western blot applications and shows reactivity with Canine, Feline, Human, Non-human primate, and Porcine 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 M3TNFABI immunogen is recombinant human TNF alpha from Pichia pastoris.
M3TNFABI 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.
Enhanced interleukin-12 and CD40 ligand activities but reduced Staphylococcus aureus Cowan 1-induced responses suggest a generalized and progressively impaired type 1 cytokine pattern for human schistosomiasis.
M3TNFABI was used in ELISA to study the role of type 1 cytokine in chronic hepatosplenic schistosomiasis
|Montenegro SM,Abath FG,Domingues AL,Melo WG,Morais CN,Coutinho EM,Mahanty S,Wynn TA||Infection and immunity (70:5903)||2002|