Detects human TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 25‑50% cross‑reactivity with recombinant porcine TNF‑ alpha and recombinant rhesus macaque TNF-alpha is observed but no cross-reactivity with recombinant cotton rat TNF‑ alpha, recombinant rat TNF‑ alpha, recombinant human (rh) LT alpha 1/ beta 2, rhLT alpha 2/ beta 1, rhAPRIL, rhBAFF, rhEDA‑A2, recombinant mouse EDA, rhFas Ligand, rhLIGHT, rhOX40 Ligand, rhTRAIL, rhTRANCE, rhTWEAK, or rhVEGI is observed.
Recombinant Monoclonal Mouse IgG1 Clone # 6401R
Protein A or G purified from cell culture supernatant
E. coli-derived recombinant human TNF-alpha Gly57-Leu233 Accession # P01375
as a solution in PBS. *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 0.01-0.06 μg/mL in the presence of 0.75 ng/mL Recombinant Human TNF-alpha and 0.5 μg/mL actinomycin D
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.
TNF‑ alpha in Human PBMCs. TNF‑ alpha was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Recombinant Mouse Anti-Human TNF‑ alpha Monoclonal Antibody (Catalog # MAB2101R) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Cytotoxicity Induced by TNF-alpha and Neutralization by Human TNF-alpha Antibody. Recombinant Human TNF-alpha (Catalog # 210-TA) induces cytotoxicity in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Human TNF-alpha (0.75 ng/mL) is neutralized (green line) by increasing concentrations of Recombinant Human TNF-alpha Monoclonal Antibody (Catalog # MAB2101R). The ND50 is typically 0.01-0.06 μg/mL in the presence of the metabolic inhibitor actinomycin D (0.5 μg/mL).
Preparation and Storage
The product is shipped with polar packs. 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 opening.
6 months, -20 to -70 °C under sterile conditions after opening.
TNF-alpha is a trimeric glycoprotein active in both membrane bound and secreted forms. TNF-alpha is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. TNF-alpha binds to 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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