The MQ2-13A5 monoclonal antibody specifically binds to human interleukin-6 (IL-6). IL-6 is a multifunctional cytokine that plays a central role in host defense mechanisms, including hematopoiesis, immune responses (eg, T and B cell activation and differentiation) and acute phase reactions. IL-6 can be expressed by a variety of cells including monocytes/macrophages, eosinophils, fibroblasts, vascular endothelial cells, bone marrow stromal cells, mesangial cells, hepatocytes, keratinocytes, astrocytes, T lymphocytes, B lymphocytes, and various tumor cells. IL-6 production is upregulated by cells in response to bacterial products such as lipopolysaccharide, viruses and other pro-inflammatory cytokines such as IL-1, TNF , and IFN-γ. IL-6 transcription is downregulated by cells in response to IL-4 and IL-10. The functional IL-6 Receptor (IL-6R) complex consists of two transmembrane glycoproteins, an 80-kDa low-affinity ligand-binding receptor subunit (IL-6Rα/CD126) and a 130 kDa (gp130/CD130) subunit that binds to IL-6-IL-6Rα to form the high-affinity signal transducing complex. Abnormal expression of IL-6 is related to the pathogenesis of many diseases including neoplastic (eg, multiple myeloma) and autoimmune diseases (eg, rheumatoid arthritis). The immunogen used to generate this hybridoma was COS-7 -expressed recombinant human IL-6.
ELISA Capture: The purified MQ2-13A5 antibody (Cat. No. 554543) is useful as a capture antibody for a sandwich ELISA for measuring human IL-6 protein levels. Purified MQ2-13A5 antibody can be paired with biotinylated MQ2-Human IL-6 39C3 antibody (Cat. No. 554546) as the detecting antibody, with recombinant human IL-6 (Cat. No. 550071) as the standard. Purified MQ2-13A5 antibody should be titrated 1-4 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of human IL-6 ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit the protocols section or chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines tested (e.g., mouse IL-1β, 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-1β, IL-2, IL-3, IL-4, IL-5, 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-1β, NT-3, PDGF-AA, sCD23 , SCF, TNF, LT-β, 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 assay of serum samples. The BD OptEIA™ ELISA Set (Cat. No. 555220) and Kit (Cat. No. 550799) are specially-formulated for serum cytokine measurement. Other Applications 1. Blocking Control for Intracellular Staining: The purified MQ2-13A5 antibody can be used as a blocking control to demonstrate specificity of IL-6 staining by the FITC-MQ2-13A5 antibody (Cat. No. 554544) or the PE-MQ2-13A5 antibody (Cat. No. 554545). To perform this control, the fixed/permeabilized cells (~ 1 million) can be incubated with 1-10 µg of purified MQ2-13A5 antibody (Cat. No. 554543) for 20 minutes at 4°C, prior to staining with the conjugated antibody (e.g., 0.1 - 0.5 µg mAb/1 million cells). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. 2. IF/Flow: The MQ2-13A5 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-6 producing cells within mixed cell populations. The directly- conjugated MQ2-13A5 antibodies are especially suitable for these studies. For specific methodology, please visit the protocols section or chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. 3. Western Blot: The MQ2-13A5 antibody (Cat. No. 554543) has been found useful for Western blot. Please note that this application is not routinely tested at BD Biosciences. 3. Neutralization: The NA/LE formulation of the MQ2-13A5 antibody (Cat. No. 554541) is useful for neutralization of human IL-6 bioactivity.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
