PE Rat Anti-Human IL-6
Clone MQ2-6A3 (RUO)
- Brand BD Pharmingen™
- Concentration 0.2 mg/ml
- Isotype Rat IgG2a, κ
- Reactivity Human (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Recombinant human IL-6
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The MQ2-6A3 antibody reacts with human interleukin-6 (IL-6). The immunogen used to generate the MQ2-6A3 hybridoma was recombinant human IL-6. This is a neutralizing antibody.
- Format PE
- Excitation Source Blue 488 nm,Green 532 nm,Yellow/Green 561 nm
- Excitation Max 496 nm
- Emission Max 578 nm
R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae. It exists in vitro as a 240-kDa protein with 23 phycoerythrobilin chromophores per molecule. This makes PE one of the brightest fluorochromes for flow cytometry applications, but its photobleaching properties make it unsuitable for fluorescence microscopy.
Suggested Companion Products
|Resources & Tools|
|Spectrum Viewer||Panel Designer||Spectrum Viewer||Download TDS||Regulatory Document Website|
Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
Immunofluorescent Staining and Flow Cytometric Analysis: The MQ2-6A3 antibody is useful for immunoflourescent staining and flow cytometric analysis to identify and enumerate human IL-6 producing cells within mixed cell populations. The PE- and FITC-conjugated MQ2-6A3 antibodies (Cat. No. 554696; 554697) are especially suitable for these experiments. (See Figure). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). 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.
A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MQ2-6A3 antibody with a ligand (e.g., recombinant human IL-6; Cat. No. 550071) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabelled MQ2-6A3 antibody prior to staining. The staining technique and blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is PE-R35-95 (Cat. No. 554689); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells).
Andersson, U. and J. Andersson. 1994. Immunolabelling of cytokine producing cells in tissues and suspension. In Cytokine Producing Cells, eds. D. Fradelizie and D. Emelie. INSERM, Paris. p. 32-49.
Assenmacher, M., J. Schmitz and A. Radbruch. 1994. Flow cytometric determination of cytokines in activated murine T helper lymphocytes: Expression of interleukin-10 in interferon-γ and in interleukin-4-expressing cells. Eur. J. Immunol. 24-1097-1101.
Austrup, F., D. Vestweber, E. Borges, M. Lohning, T. Brauer, U. Herz, H. Renz, R. Hallmann, A. Scheffold, A. Radbruch, and A.Hamann. 1997. P- and E-selectin mediate recruitment of T-helper-1 but not T-helper-2 cells into inflammed tissues. Nature. 385:81-83.
Carter, L. L., and S. L. Swain. 1997. Single cell analyses of cytokine production. Curr. Opin. Immunol. 9:177-182.
Elson, L. H., T. B. Nutman, D. D. Metcalfe and C. Prussin. 1995. Flow cytometric analysis for cytokine production identifies Thl, Th2, and Th0 cells within the human CD4+CD27- lymphocyte subpopulation. J. Immunol. 154:4294-4301.
Fujiwara, T., K. Oda, S. Yokota, S. Takatsuki and Y. Ikehara. 1988. Brefeldina A causes disassembly of the Golgi complex and accumulation of secretory proteins in the endoplasmic reticulum. J. Biol. Chem. 263:18545.
Jung, T., U. Schauer, C. Heusser, C. Neumann and C. Rieger. 1993. Detection of intracellular cytokines by flow cytometry. J. Immunol. Meth. 159:197-207.
Lecoeur, H., E. Ledru, M.-L. Gougeon. 1998. A cytofluormetric method for the simultaneous detection of both intracellular and surface antigens of apoptotic peripheral lymphocytes. J. Immunol. Meth. 217:11-26.
Parks, D. R., L. A. Herzenberg, and L. A. Herzenberg. 1989. Flow cytometry and fluorescence-activated cell sorting. In Fundamental Immunology, 2nd Edition. W. E. Paul, ed. Raven Press Ltd., New York, p. 781-802.
Picker, L. J., M. K. Singh, Z. Zdraveski, J. R. Treer, S. L. Waldrop, P. R. Bergstresser, and V. C. Maino. 1995. Direct demonstration of cytokine synthesis heterogeneity amongh human memory/effector T-cells by flow cytometry. Blood. 86:1408-1419.
Prussin, C. and D. Metcalfe. 1995. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J. Immunol. Meth. 188:117-128.
Sallusto, F., C. R. Mackay, and A. Lanzavecchia. 1997. Selective expression of the eotaxin receptor CCR3 by human T helper 2 cells. Science. 277:2005-2007.
Sander, B., I. Hoiden, U. Andersson, E. Moller, and J. Abrams. 1993. Similar frequencies and kinetics of cytokine producing cells in murine peripheral blood and spleen. J. Immunol. Meth. 166:201-214.
Sander, B., J. Andersson and U. Andersson. 1991. Assessment of cytokines by ‘immunofluorescence and the paraformalde-hyde-saponin procedure. Immunol. Rev. 119:65-93.
Vikingson, A., K. Pederson and D. Muller. 1994. Enumeration of IFN-γ producing lymphocytes by flow cytometry and correlation with quantitative measurement of IFN-γ. J. Immunol. Meth.173:219-228