Flow cytometric analysis of IL-2 expression on stimulated Rhesus macaque (Macaca mulatta) peripheral blood mononuclear cells. Rhesus PBMC were stimulated for 6 hours with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275) in the presence of 2 µM GolgiStop™ (Cat. No. 554724). The PBMC's were stained with PE Mouse Anti-Human CD3 (Cat. No. 555333/561808/561809), fixed, permeabilized, and then stained with FITC Rat Anti-Human IL-2 (0.25 µg; Cat. No. 554565/561055/559361). The quadrant markers for the bivariate dot plot was set based on the autofluorescence control, and verified with the blocking controls.
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Flow cytometric analysis of IL-2 expression on stimulated Rhesus macaque (Macaca mulatta) peripheral blood mononuclear cells. Rhesus PBMC were stimulated for 6 hours with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275) in the presence of 2 µM GolgiStop™ (Cat. No. 554724). The PBMC's were stained with PE Mouse Anti-Human CD3 (Cat. No. 555333/561808/561809), fixed, permeabilized, and then stained with FITC Rat Anti-Human IL-2 (0.25 µg; Cat. No. 554565/561055/559361). The quadrant markers for the bivariate dot plot was set based on the autofluorescence control, and verified with the blocking controls.
Product Details
BD Pharmingen™
IL2; Interleukin-2; T-cell growth factor; TCGF
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Rat IgG2a, κ
Human IL-2 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
3558
AB_2125717
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 FITC under optimum conditions, and unreacted FITC 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 mL
Cat No.
554724
FITC Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554688
FITC Rat Anti-Human IL-2 RUO
Size
25 µg
Cat No.
561055
PE Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
555333
559361 Rev. 2
Antibody Details
MQ1-17H12
The MQ1-17H12 monoclonal 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, differentiation and survival of many different target cell types including T cells, B cells, NK cells, monocytes and macrophages. The immunogen used to generate the MQ1-17H12 hybridoma was purified recombinant human IL-2 protein. The MQ1-17H12 antibody reportedly neutralizes the biological activity of human IL-2.
559361 Rev. 2
Format Details
FITC
Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
FITC
Blue 488 nm
494 nm
518 nm
559361 Rev.2
Citations & References
Development References (4)
-
Meager A. Characterization of interferons and immunoassays. In: Clemens MJ, Morris AG, Gearing AJH, ed. Lymphokines and Interferons. A Practical Approach. Oxford: IRL Press Ltd; 1987:105-127.
-
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
-
Sopper S, Stahl-Hennig C, Demuth M, Johnston IC, Dorries R, ter Meulen V. Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys. Cytometry. 1997; 29(4):351-362. (Biology). View Reference
-
Verdier F, Aujoulat M, Condevaux F, Descotes J. Determination of lymphocyte subsets and cytokine levels in cynomolgus monkeys. Toxicology. 1995; 105(1):81-90. (Biology). View Reference
559361 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.
