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Purified Mouse Anti-Human CD126
Purified 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 Purified Mouse Anti-Human CD126 (Cat. No. 551462) followed by Biotin Rat Anti-Mouse IgG1(Cat. No. 553441) and PE Streptavidin (Cat. No. 554061) in a three-layer staining protocol to amplify immunofluorescent signals. Staining with the Purified Mouse Anti-Human CD126 (filled histograms) is compared to staining obtained using Purified Mouse IgG1, κ Isotype Control (Cat. No. 555746; open histograms). Histogram in figure is gated on the CD19 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 (for example see Browning et al).
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 Purified Mouse Anti-Human CD126 (Cat. No. 551462) followed by Biotin Rat Anti-Mouse IgG1(Cat. No. 553441) and PE Streptavidin (Cat. No. 554061) in a three-layer staining protocol to amplify immunofluorescent signals. Staining with the Purified Mouse Anti-Human CD126 (filled histograms) is compared to staining obtained using Purified Mouse IgG1, κ Isotype Control (Cat. No. 555746; open histograms). Histogram in figure is gated on the CD19 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 (for example see Browning et al).
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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), ELISA Capture (Tested During Development)
0.5 mg/ml
VI C63; IX 36
3570
AB_394209
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation and Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The polyclonal antibody was purified from antiserum by affinity chromatography.

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Immunofluorescent Staining and Flow Cytometric Analysis: The Purified Mouse Anti-Human CD126 (Cat. No. 551462) can be used for the immunofluorescent staining and flow cytometric analysis of human nucleated cells to measure their expressed levels of surface IL-6Rα. An appropriate purified immunoglobulin isotype control is Purified Mouse IgG1, κ Isotype Control (Cat. No. 555746). A three-layer staining protocol is recommended for maximizing the detection of IL-6Rα expressed by cells as detailed in the figure legend.

ELISA: The Purified Mouse Anti-Human CD126 (Cat. No. 551462) can be used as the capture antibody, with recombinant soluble IL-6Ra as the standard, the Biotin Mouse Anti-Human CD126 (Cat. No. 552503) as a detection antibody for a sandwich ELISA measuring human IL-6Ra protein levels. Purified Mouse Anti-Human CD126 antibody should be titrated between 0.5-2 μg/ml to determine its optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of recombinant human soluble IL-6Ra, ranging from 500 to 2 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our website, http://www.bdbiosciences.com/us/s/resources, and go to the protocols section under "ELISA and ELISPOT".

Product Notices

  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. Ficoll-Paque is a trademark of Amersham Biosciences Limited.
  5. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551462 Rev. 4
抗体の詳細
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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.

551462 Rev. 4
フォーマットの詳細
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
551462 Rev.4
引用&参考文献
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Development References (18)

  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. Heinrich PC, Behrmann I, Müller-Newen G, Schaper F, Graeve L. Interleukin-6-type cytokine signalling through the gp130/Jak/STAT pathway. Biochem J. 1998 September; 334(Pt 2):297-314. (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 cytokine receptors by flow cytometry. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W, ed. Current Protocols in Immunology. New York: Green Publishing Associates and Wiley-Interscience; 1995:6.21.1-6.21.18.
  18. Zola H. Detection of receptors for cytokines and growth factors. Immunologist. 1994; 2:47-50. (Biology).
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551462 Rev. 4

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