Expression of IL-2 by stimulated CD3+ and CD3- human lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 18 h with PMA (Phorbol 12-Myristate 13-Acetate; Sigma, Cat. #P-8139) and Calcium Ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ Protein Transport Inhibitor (2 µM final concentration; Cat. No. 554724). The cells were fixed, permeabilized, and then stained with APC Mouse Anti-Human CD3 (Cat No. 555335) antibody and BD Horizon™ V450 Mouse Anti-Human IL-2 antibody (Cat No. 560867, Left Panel) or BD Horizon™ V450 Mouse IgG1, κ Isotype Control (Cat No.560373, Right Panel) by using BD Biosciences Pharmingen's Intracellular Cytokine Staining Protocol. The two-color flow cytometric dot plots showing the correlated expression of CD3 and IL-2 or background control staining were derived from gated events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Expression of IL-2 by stimulated CD3+ and CD3- human lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 18 h with PMA (Phorbol 12-Myristate 13-Acetate; Sigma, Cat. #P-8139) and Calcium Ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ Protein Transport Inhibitor (2 µM final concentration; Cat. No. 554724). The cells were fixed, permeabilized, and then stained with APC Mouse Anti-Human CD3 (Cat No. 555335) antibody and BD Horizon™ V450 Mouse Anti-Human IL-2 antibody (Cat No. 560867, Left Panel) or BD Horizon™ V450 Mouse IgG1, κ Isotype Control (Cat No.560373, Right Panel) by using BD Biosciences Pharmingen's Intracellular Cytokine Staining Protocol. The two-color flow cytometric dot plots showing the correlated expression of CD3 and IL-2 or background control staining were derived from gated events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Horizon™
IL2; Interleukin-2; T-cell growth factor; TCGF
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human IL-2 protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
3558
AB_10565978
Aqueous buffered solution containing protein stabilizer 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™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.
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).
- BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is manufactured and sold under license from Pestka Biomedical Laboratories, Inc. (d/b/a PBL InterferonSource) and may be used solely as indicated. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics is strictly prohibited. This product is covered by U.S. Patent No. 5,597,901 and Bulgarian Patent No. BG1895.
- All other brands are trademarks of their respective owners.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 mL
Cat No.
554724
V450 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560373
APC Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
555335
560867 Rev. 2
Antibody Details
5344.111
The 5344.111 antibody specifically binds to the multifunctional cytokine, human interleukin-2 (IL-2). IL-2 is produced by activated T cells and has multiple functions that can affect the growth, proliferation and differentiation of many different target cell types including T cells, B cells, NK cells, monocytes and macrophages. The immunogen used to generate the 5344.111 hybridoma was affinity-purified, recombinant human IL-2 protein.
The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.
560867 Rev. 2
Format Details
V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
V450
Violet 405 nm
405 nm
450 nm
560867 Rev.2
Citations & References
Development References (2)
-
Malek TR. The biology of interleukin-2. Annu Rev Immunol. 2008; 26:453-479. (Biology). View Reference
-
Waldmann TA. The biology of interleukin-2 and interleukin-15: implications for cancer therapy and vaccine design. Nat Rev Immunol. 2006; 6(8):595-601. (Biology). View Reference
560867 Rev. 2
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.
