Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified MAb1 antibody (Cat. No. 551220) is useful as a capture antibody for a sandwich ELISA for measuring human TNF protein levels. 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. The purified MAb1 antibody should be titrated between 2 - 6 µg/ml to determine its optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of TNF standard ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook.
Note: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring human TNF in serum or plasma our human TNF BD OptEIA™ ELISA Set (Cat. No. 555212) or BD OptEIA™ ELISA Kit (Cat. No. 550610) are specially formulated and recommended.
Western Blot: The MAb1 antibody has been reported to be useful for Western blot; however, the antibody is not routinely tested for this application at BD Biosciences Pharmingen.
Neutralization: The MAb1 antibody is available in the no azide/low endotoxin format (NA/LE) as Cat. No. 554508, useful for neutralization of human TNF protein bioactivity.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
The MAb1 antibody reacts with human tumor necrosis factor (TNF, aka TNF-α) 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.
Development References (2)
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Danis VA, Franic GM, Rathjen DA, Brooks PM. Effects of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-2, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and IL-6 on the production of immunoreactive IL-1 and TNF-alpha by human monocytes. Clin Exp Immunol. 1991; 85(1):143-150. (Clone-specific: ELISA). View Reference
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Rathjen DA, Cowan K, Furphy LJ, Aston R. Antigenic structure of human tumour necrosis factor: recognition of distinct regions of TNF alpha by different tumour cell receptors. Mol Immunol. 1991; 28(1-2):79-86. (Clone-specific: ELISA, Neutralization). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.