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Mouse TNF-alpha Protein (RMTNFAI) in ELISA
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 100 µL 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 1 µg/mL Actinomycin D, 2X Gentamicin and 10% Horse Serum, and 100 µL was added to each well. The plate was incubated for 23 hours at 37°C and then... View More
Product Details
RMTNFAI
Applications
Tested Dilution
Publications
Product Specifications
Species
Mouse
Published species
Not Applicable
Expression System
P. pastoris
Amino acid sequence
Mouse TNF-alpha, amino acids 80-235 (Accession # P06804)
Molecular weight
17.3 kDa
Class
Recombinant
Type
Protein
Purity
>95% by SDS-PAGE
Activity
Determined by the cytolytic effect on mouse WEHI-13VAR cells in the presence of Actinomycin D.
Conjugate
Form
Lyophilized
Purification
purified
Storage buffer
PBS
Contains
no preservative
Storage conditions
4° C
Shipping conditions
Wet ice
Product Specific Information
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 4ºC. Reconstitute in 0.5 mL of distilled or deionized water. Dilute in a buffer-containing carrier protein, aliquot, and store at or below -70ºC, avoiding multiple freeze-thaw cycles. PBS with 1% BSA or a neutral media containing <10% FCS are suitable buffers.
Target Information
TNF alpha is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophage and bind to its receptors, TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF alpha is involved in the regulation of immune cells, cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS-PAGE under non-reducing conditions. Knockout studies in mice also suggested the neuroprotective function of TNF alpha, and has been observed to causes tumor necrosis when injected into tumor-bearing mice. Other functions of TNF-alpha include its role in the immune response to bacterial, viral, parasitic and certain fungal infections, as well as its role in the necrosis of specific tumors. TNF alpha causes cytolysis or cytostasis of certain transformed cells, being synergistic with interferon-gamma in its cytotoxicity. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Bioinformatics
Protein Aliases: Cachectin; DADB-70P7.1; TNF alpha; TNF superfamily; TNF α; TNF-a; TNF-alpha; TNFα; Tumor necrosis factor; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor-alpha
Gene Aliases: DIF; Tnf; TNF-a; TNF-alpha; Tnfa; TNFalpha; Tnfsf1a; TNFSF2
UniProt ID: (Mouse) P06804
Entrez Gene ID: (Mouse) 21926
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