Detects TNF‑ alpha in ELISAs and Western blots. In sandwich immunoassays, less than 0.3% cross-reactivity with recombinant human TNF-alpha, recombinant mouse TNF-alpha, recombinant rat TNF-alpha, recombinant feline TNF-alpha, recombinant equine TNF-alpha, recombinant porcine TNF-alpha, recombinant cotton rat TNF-alpha, and rhesus macaque TNF-alpha is observed.
Polyclonal Goat IgG
E. coli-derived recombinant bovine TNF-alpha Leu78-Leu234 Accession # Q06599
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
TNF‑ alpha in Bovine PBMCs.
TNF‑ alpha was detected in immersion fixed bovine peripheral blood mononuclear cells (PBMCs) using Goat Anti-Bovine TNF‑ alpha Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF2279) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Tumor necrosis factor alpha (TNF-alpha, also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-alpha is produced by a wide variety of immune and epithelial cell types (1, 2). Bovine TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 178 aa extracellular domain (ECD) (3). Within the ECD, bovine TNF-alpha shares 64%-83% sequence identity with canine, cotton rat, equine, feline, human, mouse, porcine, rat, and rhesus TNF-alpha. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer (4). Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness (5). Cleavage of membrane bound TNF-alpha by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-alpha (6, 7). TNF-alpha trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers (1, 8). TNF-alpha regulates lymphoid tissue development through control of apoptosis (2). It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages (2). TNF-alpha is a key cytokine in several inflammatory disorders (9). It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism (10, 11).
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Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
Cludts, I. et al. (1993) Cytokine 5:336.
Tang, P. et al. (1996) Biochemistry 35:8216.
Eissner G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
Black, R.A. et al. (1997) Nature 385:729.
Moss, M.L. et al. (1997) Nature 385:733.
Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
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