The MAb11 monoclonal antibody specifically binds to human tumor necrosis factor (TNF, also known as TNF-α) protein. TNF is an efficient juxtacrine, paracrine and endocrine mediator of inflammatory and immune functions. It regulates the growth and differentiation of a variety of cell types. TNF is cytotoxic for transformed cells when in conjunction with IFN-γ. It is secreted by activated monocytes/macrophages and other cells such as B cells, T cells and fibroblasts. The immunogen used to generate the MAb11 hybridoma was recombinant human TNF. The MAb11 antibody has been reported to crossreact with Rhesus Macaque TNF.
ELISA Detection: The biotinylated MAb11 antibody (Cat. No. 554511) can be used as a detecting antibody for a sandwich ELISA for measuring human TNF protein levels. Biotinylated MAb11 antibody can be paired with the purified MAb1 antibody (Cat. No. 551220) as the capture antibody, with recombinant human TNF (Cat. No. 554618) as the standard. This detecting antibody solution should be titrated from 0.5 - 2 µg/ml to determine its optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of TNF protein ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, http://www.bdbiosciences.com/us/s/resources, and go to the protocols section under "ELISA and ELISPOT". Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines tested (e.g., mouse IL-1ß, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, GM-CSF, IFN-γ, MCP-1, TCA-3, TNF; human IL-1α, IL-1ß, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, G-CSF, GM-CSF, IFN-γ, lymphotactin, MCP-1, MCP-2, MIP-1α, MIP-1ß, NT3, PDGF-AA, sCD23 , SCF, LT-α, VEGF; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF). Note 2: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for measuring serum samples. For testing human TNF in complex biological fluids like serum or plasma, our human TNF BD OptEIA™ ELISA Set (Cat. No.555212) and BD OptEIA™ ELISA Kit II (Cat. No. 550610) are recommended. IF/Flow: The MAb11 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate TNF producing cells within mixed cell populations. FITC-conjugated MAb11 antibody (FITC-MAb11; Cat. No. 554512) and PE-conjugated MAb11 antibody (PE-MAb11; Cat. No. 554513) are recommended for these studies.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. An isotype control should be used at the same concentration as the antibody of interest.
3. 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.
4. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
