Two-color flow cytometric analysis of IL-2 expressed in stimulated human peripheral blood mononuclear cells. HiCK-1 Human Cytokine Positive Control Cells (Cat. No. 555061) were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 561811/555335/561810) and either BD Horizon™ BV510 Mouse IgG1, κ Isotype Control (Cat No. 562946; Left Panel) or with BD Horizon™ BV510 Mouse Anti-Human IL-2 antibody (Cat No. 563265; Right Panel). Two-color flow cytometric dot plots show IL-2 expression (or Ig Isotype control staining) versus CD3 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-2 expressed in stimulated human peripheral blood mononuclear cells. HiCK-1 Human Cytokine Positive Control Cells (Cat. No. 555061) were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 561811/555335/561810) and either BD Horizon™ BV510 Mouse IgG1, κ Isotype Control (Cat No. 562946; Left Panel) or with BD Horizon™ BV510 Mouse Anti-Human IL-2 antibody (Cat No. 563265; Right Panel). Two-color flow cytometric dot plots show IL-2 expression (or Ig Isotype control staining) versus CD3 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
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_2738105
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™ BV510 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV510 were 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).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Brilliant Violet™ 510 is a trademark of Sirigen.
- 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.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
BV510 Mouse IgG1, k Isotype Control RUO
Size
50 µg
Cat No.
562946
HiCK-1 Human Cytokine Positive Control Cells RUO
Size
1 mL
Cat No.
555061
APC Mouse Anti-Human CD3 RUO
Size
500 Tests
Cat No.
561811
APC Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
555335
563265 Rev. 1
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 was conjugated to BD Horizon™ BV510 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 405-nm and Em Max at 510-nm, BD Horizon™ BV510 can be excited by the violet laser and detected in the BD Horizon™ V500 (525/50-nm) filter set. BD Horizon™ BV510 conjugates are useful for the detection of dim markers off the violet laser.
563265 Rev. 1
Format Details
BV510
The BD Horizon Brilliant Violet™ 510 (BV510) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye with an excitation maximum (Ex Max) at 327-nm / 405-nm and an emission maximum (Em Max) at 512-nm. BV510, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 510-nm (e.g., a 525/50 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.
BV510
Violet 405 nm
327 nm, 405 nm
512 nm
563265 Rev.1
Citations & References
Development References (4)
-
Liao W, Lin JX, Leonard WJ. IL-2 family cytokines: new insights into the complex roles of IL-2 as a broad regulator of T helper cell differentiation. Curr Protoc Immunol. 2011; 23(5):598-604. (Biology). View Reference
-
Malek TR. The biology of interleukin-2. Annu Rev Immunol. 2008; 26:453-479. (Biology). View Reference
-
Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology). 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
563265 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.
