ELISA analysis of TNF-alpha secreted from ovine AH-1 trophoblasts at various time points after infection with Chlamydia abortus (at a multiplicity of infection (MOI) = 1) was performed by coating 100ul per well of anti-bovine TNF-alpha polyclonal antibody (Product # PBOTNFAI) at a concentration of 10ug/ml and incubating overnight at 4C. The plate was blocked for one hour using 3% bovine serum albumin (BSA) diluted in PBST. The plate was washed six times with 200ul/well PBST, and then incubated with 100ul per well of tissue culture medium from uninfected AH-1 cells (negative control) or AH-1 cells infected with Chlamydia abortus, for one hour at room temperature. The plate was washed with wash buffer six times and incubated with 100ul per well anti-bovine TNF-alpha biotin-conjugated polyclonal antibody (Product # PBOTNFABI) at 1ug/ml for one hour at room temperature. The plate was washed with wash buffer six times and then the signal was amplified using HRP-conjugated strepavidin at 2ug/ml for one hour at room temperature. Detection was performed using 100ul per well 3,3',5,5'-tetramethylbenzidine (TMB) for 10 minutes. The reaction was stopped with 0.2M sulfuric acid. Absorbance in optical density (OD) values were read on a spectrophotometer at 450nm. Data courtesy of the Innovators Program.
|Tested species reactivity||Bovine, Sheep|
|Host / Isotype||Rabbit / IgG|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|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.
PBOTNFABI targets TNF Alpha in ELISA, and WB applications and shows reactivity with bovine and ovine samples.
The PBOTNFABI immunogen is tNF alpha.
PBOTNFABI detects TNF Alpha which has a predicted molecular weight of approximately 26 kDa.
The PBOTNFABI polyclonal biotinylated bovine TNFa antibody has successfully been paired as the detection antibody in a sandwich ELISA with monoclonal capture antibody MBOTNFAI (clone E10011). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
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.