Detects rabbit TNF-alpha in direct ELISAs.
In direct ELISAs, no cross-reactivity
with recombinant TNF-alpha from cotton rat, cat, cow, dog, guinea pig, horse,
human, mouse, pig, rat, or rhesus is observed. No cross-reactivity with
recombinant human (rh) 4-1BB Ligand, rhAPRIL, rhBAFF, rhEDA, rhEDA-A2, rhFas
Ligand, rhGITR Ligand, rhLIGHT, rhLymphotoxin alpha 1 beta 2, rhLymphotoxin alpha 2 beta 1, rhOX40
Ligand, rhTRAIL, rhTRANCE, rhTWEAK, rhVEGI, recombinant mouse (rm) 4-1BB
Ligand, rmBAFF, rmFas Ligand, rmGITR Ligand, rmLIGHT, rmOX40 Ligand, rmTRAIL,
rmTRAIL-2L, rmTRANCE, or rmTWEAK is observed.
Monoclonal Mouse IgG2A Clone # 610331
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant rabbit TNF-alpha Leu79-Leu234 Accession # NP_001075732
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. Matthews, N. and M. L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M. J. et al. (eds): IRL Press. 221. The Neutralization Dose (ND50) is typically 0.4-2.4 µg/mL in the presence of 0.25 ng/mL Recombinant Rabbit TNF‑ alpha and the metabolic inhibitor actinomycin D (1 µg/mL).
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Cytotoxicity Induced by TNF‑ alpha and Neutralization by Rabbit TNF‑ alpha Antibody. Recombinant Rabbit TNF‑ alpha (Catalog # 5670-TG) induces cytotoxicity in the the L‑929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin. Cytotoxicity elicited by Recombinant Rabbit TNF‑ alpha (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Rabbit TNF‑ alpha Monoclonal Antibody (Catalog # MAB5670). The ND50 is typically 0.4‑2.4 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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 (TNF-alpha ), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily and family. The 26 kDa type II transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Rabbit TNF-alpha is 235 amino acids (aa) in length and contains a 35 aa cytoplasmic domain, a 21 aa transmembrane region, and a 179 extracellular domain (ECD). Cleavage of membrane bound TNF-alpha by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-alpha. Rabbit TNF-alpha is 79% and 78% aa identical to human and mouse TNF-alpha, respectively. TNF-alpha is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. TNF-alpha binds TNF RI and TNF RII present on virtually all cell types where it triggers the activation of multiple signal transduction pathways and modulates the expression of a wide variety of genes.
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