Neutralization Activity: This antibody has been reported to be useful for the neutralization of recombinant human TNF. Neutralization activity may be measured using a cytolysis assay for MTT dye conversion with 2.0 ng/mL recombinant human TNF (Cat. No. 554618) and L929 at 2x10^5 cells/mL as indicator cells (i.e. preincubation of the antibody with recombinant human TNF for 60 minutes). 50% Neutralization (ND50) at 20 - 1200 ng/mL > 85% Neutralization at 2 - 15 µg/mL
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Sterility
0.2 μm filtered
Endotoxin Level
Endotoxin level is ≤ 0.01 EU/μg (≤ 0.001 ng/μg) of protein as determined by the LAL assay.
ELISA: The purified format of the MAb1 antibody (Cat. No. 551220) is useful as a capture antibody for a sandwich ELISA for measuring human TNF. The purified MAb1 antibody can be paired with the biotinylated MAb11 antibody (Cat. No. 554511) as the detection antibody, with recombinant human TNF protein (Cat. No. 554618) as the standard. For complex biological samples, such as plasma or serum, investigators are encouraged to consider utilizing the BD OptEIA™ ELISA Set (Cat. No. 555212) or BD OptEIA™ ELISA Kit (Cat. No. 550610).
Restrictions
For Research Use only
Format
Liquid
Concentration
1.0 mg/mL
Buffer
No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2μm sterile filtered.
Preservative
Azide free
Storage
4 °C
Storage Comment
Store undiluted at 4°C. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
Rathjen, Cowan, Furphy, Aston: "Antigenic structure of human tumour necrosis factor: recognition of distinct regions of TNF alpha by different tumour cell receptors." in: Molecular immunology, Vol. 28, Issue 1-2, pp. 79-86, (1991) (PubMed).
Hogan, Vogel: "Production of tumor necrosis factor by rIFN-gamma-primed C3H/HeJ (Lpsd) macrophages requires the presence of lipid A-associated proteins." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 141, Issue 12, pp. 4196-202, (1989) (PubMed).
The MAb1 antibody reacts with human tumor necrosis factor (TNF, aka TNF-alpha) protein. TNF is a multifunctional cytokine involved in a variety of immune and inflammatory responses including hemorrhagic necrosis of tumors, septic shock, fever and autoimmune diseases. TNF can regulate the growth and differentiation of many different cell types. It is produced by different activated cell types including macrophages, T lymphocytes, NK cells, dendritic cells, endothelial cells, peripheral blood leukocytes, osteoblasts, astrocytes, mast cells, Kupffer cells, smooth muscle cells, fibroblasts and certain tumor cells. TNF exists in two biologically active forms, i.e., transmembrane and soluble forms. Upon activation, cells express transmembrane TNF glycoproteins that associate as homotrimeric complexes. After enzymatic cleavage, the extracellular region of membrane TNF sheds as a soluble homotrimer. The biologically active form of TNF has been reported to be a trimer.