Phone:
+1 877 302 8632
Fax:
+1 888 205 9894 (Toll-free)
E-Mail:
orders@antibodies-online.com

IL-6 Receptor antibody (PE-Cy7)

IL6R Reactivity: Human FACS Host: Mouse Monoclonal UV4 PE-Cy7
Catalog No. ABIN2659081
  • Target See all IL-6 Receptor (IL6R) Antibodies
    IL-6 Receptor (IL6R) (Interleukin 6 Receptor (IL6R))
    Reactivity
    • 49
    • 19
    • 16
    Human
    Host
    • 47
    • 22
    • 5
    • 1
    • 1
    Mouse
    Clonality
    • 50
    • 25
    • 1
    Monoclonal
    Conjugate
    • 49
    • 13
    • 5
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This IL-6 Receptor antibody is conjugated to PE-Cy7
    Application
    • 51
    • 23
    • 22
    • 21
    • 12
    • 9
    • 4
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    Flow Cytometry (FACS)
    Purification
    The antibody was purified by affinity chromatography and conjugated with PE/Cy7 under optimal conditions. The solution is free of unconjugated PE/Cy7 and unconjugated antibody.
    Clone
    UV4
    Isotype
    IgG1 kappa
    Top Product
    Discover our top product IL6R Primary Antibody
  • Application Notes
    Optimal working dilution should be determined by the investigator.
    Restrictions
    For Research Use only
  • Validation #101960 (Flow Cytometry)
    'Independent Validation' Badge
    by
    Immunmodulatorische Abteilung, Universitätsklinikum Erlangen
    No.
    #101960
    Date
    11/06/2017
    Antigen
    IL6R
    Lot Number
    B236171
    Method validated
    Flow Cytometry
    Positive Control
    Mature dendritic cells (mDCs) from three donors, incubated with ABIN2659082
    Negative Control
    Mature dendritic cells (mDCs) from three donors, incubated without antibody
    Notes

    Passed. ABIN2659082 specifically recognizes the IL-6 Receptor on mDCs.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN2659082
    Secondary Antibody
    Full Protocol
    • Prepare peripheral blood mononuclear cells (PBMCs) from leukoreduction system chambers (LRSCs) (Pfeiffer et al. (2013); Weidinger et al. 2011) from three healthy donors (obtained following informed consent and approved by the institutional review board) by density centrifugation on a Lymphoprep gradient (Axis-shield AS, 1114547, lot 12IFS10).
    • For the generation of dendritic cells (DCs), prepare PBMCs from leukoreduction system chambers (LRSC) (Pfeiffer et al. (2013)).
    • After centrifugation, collect PBMCs forming a distinct band at the sample/Lymphoprep interface.
    • Wash PBMCs 3x with cold PBS supplemented with 1mM EDTA.
    • Wash PBMCs 1x with cold RPMI 1640 (Lonza, BE12-167F, lot 7MB088).
    • Resuspend PBMCs in warm DC Medium (RPMI 1640 supplemented with 1% AB-serum, 100U/ml penicillin, 100mg/ml streptomycin, 2mM L-glutamine, 10mM HEPES buffer (Lonza, BE17-737E)) and seeded into 175cm2 tissue culture flasks at a density of 3.5x108 cells/flask.
    • Let monocytes adhere for 1h at 37°C and 5% CO2.
    • Wash non-adherent fraction off with warm RPMI 1640.
    • Add 30ml DC medium supplemented with 800U/ml recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) and 250U/ml recombinant human interleukin-4 (IL-4) to each flask (day 1).
    • Incubate cells for 72h at 37°C and 5% CO2.
    • On day 4 (or three days later), add 5ml of fresh culture medium containing GM-CSF (final concentration of 400 U/ml in 35ml) and IL-4 (final concentration of 250U/ml in 35ml) to the cells.
    • On day 5, mature the immature DCs by adding 10ng/ml recombinant human tumor necrosis factor alpha (TNF-α), 1μg/ml prostaglandin E2 (PGE2), 200U/ml recombinant human interleukin-1β (IL-1β), 1000U/ml recombinant human interleukin-6 (IL-6), 40U/ml GM-CSF, and 250 U/ml IL-4 to the medium.
    • Incubate cells for 48h at 37°C and 5% CO2.
    • On day 7, use the now mature DCs (mDCs) for FACS analysis.
    • Analyze cell surface expression of IL-6 Receptor by FACS:
      • Harvest 2.5x105 mDCs by centrifugation.
      • Wash cells once with FACS buffer (PBS supplemented with 2% FCS).
      • Combine anti-human PE-Cy7 conjugated IL-6 Receptor antibody (antibodies-online, ABIN2659082, lot B236171) diluted 1:100 and LIVE/DEAD Violet dead cell stain kit (ThermoFisher Scientific, L34955, lot 1832693) diluted 1:300 in cold FACS buffer.
      • Resuspend mDCs in 100µl of the antibody-solution and incubate for 30min on ice in the dark.
      • Wash mDCs 2x with cold FACS buffer.
      • Fix mDCs using 2% paraformaldehyde in cold PBS.
      • Store mDCs at 4°C until analysis.
      • Run samples using a BD FACS Canto II and analyze events using FCS Express 5.
    Experimental Notes

    Mature DCs were stained for FACS for the analysis of the expression of IL-6 Receptor/CD126 on the cell surface of the cells with ABIN2659082 for 30min or 120min respectively. The histogram shows a clearly shift between unstained control and the stained mock control by staining for 120min. The staining for 30min does also show a shift of the histogram peak, but not as clear as the staining for 120min.

  • Buffer
    Phosphate-buffered solution, pH 7.2, containing 0.09 % sodium azide and 0.2 % (w/v) BSA .
    Preservative
    Sodium azide
    Precaution of Use
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Handling Advice
    Protect from prolonged exposure to light. Do not freeze.
    Storage
    4 °C
    Storage Comment
    The antibody solution should be stored undiluted between 2°C and 8°C.
  • Target
    IL-6 Receptor (IL6R) (Interleukin 6 Receptor (IL6R))
    Alternative Name
    CD126 (IL6R Products)
    Synonyms
    IL6R antibody, il-6ra antibody, CD126 antibody, IL-6R-1 antibody, IL-6RA antibody, IL6Q antibody, IL6RA antibody, IL6RQ antibody, gp80 antibody, IL6R1 antibody, Il6ra antibody, IL-6Ralpha antibody, IL-6R antibody, Il6r antibody, interleukin 6 receptor antibody, interleukin 6 receptor, alpha antibody, IL6R antibody, il6r antibody, Il6r antibody, Il6ra antibody
    Background
    CD126 is an 80 kD IL-6 receptor α chain also known as IL-6R. It is a member of the immunoglobulin superfamily that is expressed on plasma cells, T cells, activated B cells, monocytes, granulocytes, hepatocytes, epithelial cells, and fibroblasts. Functional IL-6 receptors are formed by the non-covalent association of CD126 and the IL-6 receptor β chain (CD130 or gp130). CD126 binds IL-6 with low affinity but does not signal. The β chain (gp130, CD130) does not bind IL-6 by itself but associates with the α-chain/IL-6 complex to initiate signal transduction. IL-6 binding to the receptor complex results in the stimulation of B and T cells, and hematopoietic precursor proliferation and differentiation. A soluble form of CD126 has been found in human serum.
    Pathways
    JAK-STAT Signaling, Autophagy, Growth Factor Binding, Cancer Immune Checkpoints
You are here:
Support