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    Alexa Fluor® 647 Mouse anti-Human IL-21

    BD Pharmingen™ Alexa Fluor® 647 Mouse anti-Human IL-21

    Clone 3A3-N2.1 (also known as 3A3-N2)

    (RUO)
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    Alexa Fluor® 647 Mouse anti-Human IL-21
    Flow cytometric analysis of Alexa 647 anti-human IL-21 on stimulated and non-stimulated PBMC. Human PBMC were cultured with media alone (left panel)  or stimulated with PMA/Ionomycin in the presence of BD GolgiStop™ (Cat. No. 554724) for 5 hours (right panel). Cells were then fixed with BD Cytofix™  Fixation Buffer (Cat. No. 554655), and stored in BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) at 4°C overnight. The cells were then permeabilized using BD Perm/Wash™ Buffer (Cat. No. 554723) followed by staining with FITC Mouse Anti-Human CD4 (clone RPA-T4, Cat. No. 555346) and Alexa 647 Mouse anti-Human IL-21. The dot plots were derived from gated lymphocytes. Flow cytometry was performed on a BD FACSCalibur™ Flow Cytometer.
    Alexa Fluor® 647 Mouse anti-Human IL-21
    Flow cytometric analysis of Alexa 647 anti-human IL-21 on stimulated and non-stimulated PBMC. Human PBMC were cultured with media alone (left panel)  or stimulated with PMA/Ionomycin in the presence of BD GolgiStop™ (Cat. No. 554724) for 5 hours (right panel). Cells were then fixed with BD Cytofix™  Fixation Buffer (Cat. No. 554655), and stored in BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) at 4°C overnight. The cells were then permeabilized using BD Perm/Wash™ Buffer (Cat. No. 554723) followed by staining with FITC Mouse Anti-Human CD4 (clone RPA-T4, Cat. No. 555346) and Alexa 647 Mouse anti-Human IL-21. The dot plots were derived from gated lymphocytes. Flow cytometry was performed on a BD FACSCalibur™ Flow Cytometer.
    Product Details
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    BD Pharmingen™
    Interleukin-21, IL21, Za11
    Human (QC Testing)
    Mouse IgG1, κ
    Human IL-21 Recombinant Protein
    Intracellular staining (flow cytometry) (Routinely Tested)
    20 µl
    AB_1645421
    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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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    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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
    3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
    5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
    6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
    7. 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.
    8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
    9. An isotype control should be used at the same concentration as the antibody of interest.
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    562043 Rev. 1
    Antibody Details
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    3A3-N2.1

    Human Interleukin-21 (IL-21) is a member of the type I cytokine family that is encoded by a gene resident on chromosome 4. The mature form of human IL-21 is a 131 amino acid protein. IL-21 is produced by activated NKT and multiple CD4+ T cell subsets including effector memory and central memory CD4+ T cells and differentiated T helper cell subsets polarized towards Th17 cell and T follicular helper (Tfh) phenotypes. IL-21 plays important protective roles in the regulation of hematopoiesis and both innate and adaptive immune responses and adverse roles in promoting autoimmunity. IL-21 costimulates the proliferation and differentiation of CD4+ T cells. It enhances the proliferation of and cytotoxicity mediated by natural killer (NK) cells and CD8+ T cells. IL-21 costimulates B cell proliferation and differentiation into plasma cells producing immunoglobulins with IgG isotypes. IL-21 can also regulate the functions of dendritic cells and other myeloid cells. IL-21 exerts its biological activities by binding to and activating the Janus activating kinases (JAK1 and JAK3) and signal transducers and activators of transcription (STAT1, STAT3, STA5a and STAT5b) signaling pathways through the IL-21 receptor (IL-21R) complex. The IL-21R complex is comprised of the IL-21R alpha subunit and the common cytokine receptor gamma subunit (γ c; CD132).

    The monoclonal 3A3-N2 antibody specifically binds to human IL-21.

    562043 Rev. 1
    Format Details
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    Alexa Fluor™ 647
    Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
    Alexa Fluor™ 647
    Red 627-640 nm
    653 nm
    669 nm
    562043 Rev.1
    Citations & References
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    Development References (5)

    1. Coquet JM, Kyparissoudis K, Pellicci DG, et al. IL-21 is produced by NKT cells and modulates NKT cell activation and cytokine production. J Immunol. 2007; 178(5):2827-2834. (Biology). View Reference
    2. Onoda T, Rahman M, Nara H, et al. Human CD4+ central and effector memory T cells produce IL-21: effect on cytokine-driven proliferation of CD4+ T cell subsets. Int Immunol. 2007; 19(10):1191-1199. (Biology). View Reference
    3. Parrish-Novak J, Dillon SR, Nelson A, et al. Interleukin 21 and its receptor are involved in NK cell expansion and regulation of lymphocyte function. Nature. 2000; 408(6808):57-63. (Biology). View Reference
    4. Pene J, Gauchat JF, Lecart S, et al. Cutting edge: IL-21 is a switch factor for the production of IgG1 and IgG3 by human B cells. J Immunol. 2004; 172(9):5154-5157. (Biology). View Reference
    5. Spolski R, Leonard WJ. Interleukin-21: basic biology and implications for cancer and autoimmunity. Annu Rev Immunol. 2008; 26:57-79. (Biology). View Reference
    View All (5) View Less
    562043 Rev. 1

    Please refer to Support Documents for Quality Certificates


    Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


    Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

    For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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