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BV421 Mouse Anti-Human IL-21
BV421 Mouse Anti-Human IL-21

Two-color flow cytometric analysis of IL-21 expression in stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (Sigma P-8139; 50 ng/ml final concentration) and Ionomycin (Sigma I-0634; 1 μg/ml final concentration) in the presence of  BD GolgiStop™  Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655).

              The cells were then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Mouse Anti-Human CD4 antibody (Cat.No. 561841/555349/561840) and either BD Horizon™ BV421 Mouse IgG1 κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse Anti-Human IL-21 antibody (Cat. No. 564755; Right Panel) using the BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression of IL-21 (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Two-color flow cytometric analysis of IL-21 expression in stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (Sigma P-8139; 50 ng/ml final concentration) and Ionomycin (Sigma I-0634; 1 μg/ml final concentration) in the presence of  BD GolgiStop™  Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655).

              The cells were then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Mouse Anti-Human CD4 antibody (Cat.No. 561841/555349/561840) and either BD Horizon™ BV421 Mouse IgG1 κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse Anti-Human IL-21 antibody (Cat. No. 564755; Right Panel) using the BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression of IL-21 (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
Interleukin-21, IL21, Za11
Human (QC Testing)
Mouse IgG1, κ
Human IL-21 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738933
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  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.
564755 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.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

564755 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
564755 Rev.1
Citations & References
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Development References (7)

  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. Lindqvist M, van Lunzen J, Soghoian DZ, et al. Expansion of HIV-specific T follicular helper cells in chronic HIV infection. J Clin Invest. 2012; 122(9):3271-3280. (Clone-specific: Flow cytometry). View Reference
  3. 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
  4. 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
  5. 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
  6. Schmitt N, Morita R, Bourdery L, et al. Human dendritic cells induce the differentiation of interleukin-21-producing T follicular helper-like cells through interleukin-12. Immunity. 2009; 31(1):158-169. (Clone-specific: Flow cytometry). View Reference
  7. 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 (7) View Less
564755 Rev. 1

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