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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. 561656/551850; filled histogram) or PE Mouse IgG1, κ Isotype Control (Cat. No 555749; open histogram). 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.
BD Pharmingen™ PE Mouse Anti-Human CD126
Regulatory Status Legend
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
Recommended Assay Procedures
Immunofluorescent Staining and Flow Cytometric Analysis: The PE Mouse Anti-Human CD126 (Cat. No. 561696/551850) antibody can be used for immunofluorescent staining (20 µg/10e6 cells) and flow cytometric analysis of human nucleated cells to measure their expressed levels of surface IL-6Rα. An appropriate immunoglobulin isotype control is PE Mouse IgG1, κ Isotype Control (Cat. No. 555749).
ELISA: The Purified Mouse Anti-Human CD126 antibody (Cat. No. 551462) is useful as a capture for a sandwich ELISA that measures soluble human IL-6Rα protein levels. The M5 antibody can be paired with the M182 antibody and recombinant soluble human IL-6Rα as a standard.
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.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Ficoll-Paque is a trademark of Amersham Biosciences Limited.
- 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 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.
Development References (17)
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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
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