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PE Mouse Anti-Human CD126
PE Mouse Anti-Human CD126

Expression of cell surface IL-6R by human PBMC. Human PBMC isolated by density centrifugation (Ficoll-Paque™) were blocked with normal polyclonal human IgG and stained with PE Mouse Anti-Human CD126 (Cat No. 551850). Staining with PE Mouse Anti-Human CD126  (filled histograms) is compared to staining obtained using PE Mouse IgG1, κ Isotype Control (Cat. No. 555749; open histograms). Histograms in the figure were gated on the CD19-negative lymphocytes. Note: Certain human cell lines or cell types (e.g., neutrophils, monocytes) can first be treated with reagents that block receptors for the Fc regions of immunoglobulin to avoid nonspecific immunofluorescent staining mediated by Fc receptors.

Expression of cell surface IL-6R by human PBMC. Human PBMC isolated by density centrifugation (Ficoll-Paque™) were blocked with normal polyclonal human IgG and stained with PE Mouse Anti-Human CD126 (Cat No. 551850). Staining with PE Mouse Anti-Human CD126  (filled histograms) is compared to staining obtained using PE Mouse IgG1, κ Isotype Control (Cat. No. 555749; open histograms). Histograms in the figure were gated on the CD19-negative lymphocytes. Note: Certain human cell lines or cell types (e.g., neutrophils, monocytes) can first be treated with reagents that block receptors for the Fc regions of immunoglobulin to avoid nonspecific immunofluorescent staining mediated by Fc receptors.

Product Details
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BD Pharmingen™
Interleukin 6 Receptor alpha chain; IL-6R alpha; IL-6Rα
Human (QC Testing)
Mouse BALB/c IgG1, κ
CD126 Recombinant Protein
Flow cytometry (Routinely Tested)
20 µl
VI C63; IX 36
3570
AB_394271
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

Immunofluorescent Staining and Flow Cytometric Analysis: The PE Mouse Anti-Human CD126 (Cat. No. 551850) can be used for the immunofluorescent staining (20 µg/10e6 cells) and flow cytometric analysis of human nucleated cells to measure their expressed levels of surface IL-6Ra.15 An appropriate purified immunoglobulin is PE Mouse IgG1, κ Isotype Control (Cat. No. 555749).

ELISA: Purified Mouse Anti-Human CD126 (Cat. No. 551462) is useful as a capture for a sandwich ELISA that measures soluble human IL-6Ra protein levels. The M5 antibody can be paired with the M182 antibody and recombinant soluble human IL-6Ra as a standard.

Product Notices

  1. 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).
  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. Ficoll-Paque is a trademark of Amersham Biosciences Limited.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551850 Rev. 2
Antibody Details
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M5

The M5 monoclonal antibody specifically binds to human CD126 which is also known as the alpha subunit of the human IL-6 Receptor (IL-6Rα). CD126 is an 80 kDa type I transmembrane glycoprotein, also known as gp80 and B cell stimulatory factor-2 (BSF-2) Receptor. The IL-6Rα subunit associates with the 130-160 kDa gp130 subunit (IL-6 Receptor β chain, CD130), that is shared with the receptor complexes for Leukemia Inhibitory Factor (LIF), Ciliary Neurotropic Factor (CNTF), Oncostatin M (OSM), IL-11, Cardiotropin 1 (CT-1) and possibly Neurotrophin-1/B Cell-Stimulating Factor 3 (NNT-1/BSF-3).  The IL-6Rα chain binds IL-6 with low affinity; however the association with CD130 stabilizes the IL-6/IL-6Rα complex resulting in the formation of a high affinity ligand-receptor complex. The IL-6Rβ chain mediates signal transduction. CD126 is expressed at high levels by activated and EBV-transformed B cells, plasma cells and myeloma cells and at lower levels by most leucocytes, epithelial cells, fibroblasts, hepatocytes and neural cells. IL-6Rα exists in soluble form in human serum. The serum levels of soluble IL-6Rα appear to elevate in pathological situations such as multiple myeloma, Grave's disease, juvenile chronic arthritis and HIV. The M5 antibody is directed against an epitope not involved in interactions of CD126 with IL-6 or CD130.

551850 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
551850 Rev.2
Citations & References
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Development References (17)

  1. Browning JL, Dougas I, Ngam-ek A, et al. Characterization of surface lymphotoxin forms. Use of specific monoclonal antibodies and soluble receptors.. J Immunol. 1995; 154(1):33-46. (Biology). View Reference
  2. Gaillard JP, Bataille R, Brailly H, et al. Increased and highly stable levels of functional soluble interleukin-6 receptor in sera of patients with monoclonal gammopathy. Eur J Immunol. 1993 April; 23(4):820-824. (Biology). View Reference
  3. Gaillard JP, Mani JC, Liautard J, Klein B, Brochier J. Interleukin-6 receptor signaling. I. gp80 and gp130 receptor interaction in the absence of interleukin-6.. Eur Cytokine Netw. 1999; 10(1):43-8. (Biology). View Reference
  4. Hibi M, Murakami M, Saito M, Hirano T, Taga T, Kishimoto T. Molecular cloning and expression of an IL-6 signal transducer, gp130. Cell. 1990; 63(6):1149-1157. (Biology). View Reference
  5. Hirano T, Nakajima K, Hibi M. Signaling mechanisms through gp130: a model of the cytokine system. Cytokine Growth Factor Rev. 1997 December; 8(4):241-252. (Biology). View Reference
  6. Honda M, Yamamoto S, Cheng M, et al. Human soluble IL-6 receptor: its detection and enhanced release by HIV infection. J Immunol. 1992 April; 148(7):2175-2180. (Biology). View Reference
  7. Keul R, Heinrich PC, Müller-newen G, Muller K, Woo P. A possible role for soluble IL-6 receptor in the pathogenesis of systemic onset juvenile chronic arthritis. Cytokine. 1998 September; 10(9):729-734. (Biology). View Reference
  8. Liautard J, Gaillard JP, Mani JC, et al. Epitope analysis of human IL-6 receptor gp80 molecule with monoclonal antibodies.. Eur Cytokine Netw. 1994 May-June; 5(3):293-300. (Biology). View Reference
  9. Llinas L, Lazaro A, de Salort J, Matesanz-Isabel J, Sintes J, Engel P. Expression profiles of novel cell surface molecules on B-cell subsets and plasma cells as analyzed by flow cytometry. Immunol Lett. 2011; 134(2):113-121. (Biology). View Reference
  10. Müller-Newen G, Köhne C, Keul R, et al. Purification and characterization of the soluble interleukin-6 receptor from human plasma and identification of an isoform generated through alternative splicing. Eur J Biochem. 1996 March; 236(3):837-842. (Biology). View Reference
  11. Salvi M, Girasole G, Pedrazzoni M, et al. Increased serum concentrations of interleukin-6 (IL-6) and soluble IL-6 receptor in patients with Graves' disease. J Clin Immunol. 1996 August; 81(8):2976-2979. (Biology). View Reference
  12. Senaldi G, Varnum BC, Sarmiento U, et al. Novel neurotrophin-1/B cell-stimulating factor-3: a cytokine of the IL-6 family. Proc Natl Acad Sci U S A. 1999 September; 96(20):11458-11463. (Biology). View Reference
  13. Taga T, Hibi M, Hirata Y, et al. Interleukin-6 triggers the association of its receptor with a possible signal transducer, gp130. Cell. 1989 August; 58(3):573-581. (Biology). View Reference
  14. Taga T, Kawanishi Y, Hardy RR, Hirano T, Kishimoto T. Receptors for B cell stimulatory factor 2. Quantitation, specificity, distribution, and regulation of their expression. J Exp Med. 1987 October; 166(4):967-981. (Biology). View Reference
  15. Van Snick J. Interleukin-6: an overview. Annu Rev Immunol. 1990; 8:253-278. (Biology). View Reference
  16. Yamasaki K, Taga T, Hirata Y, et al. Cloning and expression of the human interleukin-6 (BSF-2/IFN beta 2) receptor. Science. 1988 August; 241(4867):825-828. (Biology). View Reference
  17. Zola H. Detection of receptors for cytokines and growth factors. Immunologist. 1994; 2:47-50. (Clone-specific: Flow cytometry).
View All (17) View Less
551850 Rev. 2

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