Cytotoxicity Bioassay Analysis of Murine TNF-alpha was performed by resuspending 929 cells in 10% Horse Serum. Six thousand cells per well (passage 8, in 100ul volume) were transferred to assay plate and incubated overnight at 37°C. Mouse TNF-alpha Recombinant Protein (Product # RMTNFAI), was serially diluted 1:4 in triplicate starting from a concentration of 100 ng/mL in assay media containing 1ug/mL Actinomycin D, 2X Gentamicin and 10% Horse Serum, and 100ul was added to each well. The plate was incubated for 23 hours at 37°C and then analyzed via a colorimetric method to determine cell viability and read at 490nm.
|Tested species reactivity||Mouse|
|Host / Isotype||Yeast|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Control (Ctrl)||Assay Dependent|
|ELISA (ELISA)||Assay Dependent|
|Immunoprecipitation (IP)||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.
The mature, soluble form of mouse TNF-alpha was expressed in Pichia pastoris, KM71H. GenBank Accession # P06804, amino acids 80 - 235. For protein verification, the expressed protein was N-terminal sequenced and begins with the following 10 amino acids: LRSSSQNSSD. Purity is >95% by SDS-PAGE analyses.
Activity: The biological activity of recombinant mouse TNFa was measured by its cytolytic effect on mouse WEHI-13VAR cells in the presence of Actinomycin D.
Use: For in vitro applications. A dose-response assay should be performed to determine the optimal concentration for specific experiments.
Reconstitution and Dilution: Store lyophilized material at 4°C. Reconstitute in 0.5 ml of distilled or deionized water. Dilute in a buffer-containing carrier protein, aliquot, and store at or below -70°C, avoiding multiple freeze-thaw cycles. PBS with 1% BSA or a neutral media containing <10% FCS are suitable buffers.
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.