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Flow cytometric analysis of IL-6 expression by stimulated human monocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 6 hours with lipopolysaccharide (LPS; 100 ng/ml final concentration) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Containing Monensin) (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with PE-Mouse Anti-Human CD14 monoclonal antibody (Cat. No. 555398), fixed and permeabilized using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Perm/Wash™ Buffer (Cat. No. 554723), and stained with APC Rat Anti-Human IL-6 antibody (Cat. No. 561441, solid line histogram) or an APC Rat IgG1, κ Isotype Control (Cat No. 554686, dashed line histogram). The fluorescence histograms were derived from CD14-positive events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
BD Pharmingen™ APC Rat Anti-Human IL-6
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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.
Development References (4)
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Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Biology: ELISA, Neutralization). View Reference
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Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Biology: ELISA, Neutralization). View Reference
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Gaines Das RE, Poole S. The international standard for interleukin-6. Evaluation in an international collaborative study. J Immunol Methods. 1993; 160(2):147-153. (Biology: ELISA, Neutralization). View Reference
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Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: IC/FCM Block). View Reference
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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.