Flow cytometric analysis of IL-2 expression on PMA+ CA++ ionophore-stimulated peripheral blood lymphocytes of Rhesus macaque (Macaca mulatta). PBMCs were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. #P-8139) and calcium ionophore A23187 (500-100 ng/ml final concentration; Sigma, Cat. #C-9275) in the presence of BD GolgiStop™ (2 μM final concentration; Cat. No. 554724). The PBMCs were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555916), fixed, permeabilized, and then stained with APC Rat Anti-Human IL-2 (Cat. No. 551383). While the frequency of CD3+ non-human primate cells reacting with MQ1-17H12 following PMA + Ca++ ionophore activation is similar to that observed with normal human donor CD3+ lymphocytes, the fluorescence intensity is characteristically weaker.
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Flow cytometric analysis of IL-2 expression on PMA+ CA++ ionophore-stimulated peripheral blood lymphocytes of Rhesus macaque (Macaca mulatta). PBMCs were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. #P-8139) and calcium ionophore A23187 (500-100 ng/ml final concentration; Sigma, Cat. #C-9275) in the presence of BD GolgiStop™ (2 μM final concentration; Cat. No. 554724). The PBMCs were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555916), fixed, permeabilized, and then stained with APC Rat Anti-Human IL-2 (Cat. No. 551383). While the frequency of CD3+ non-human primate cells reacting with MQ1-17H12 following PMA + Ca++ ionophore activation is similar to that observed with normal human donor CD3+ lymphocytes, the fluorescence intensity is characteristically weaker.
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_398503
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 APC under optimum conditions, and unconjugated antibody and free APC 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- 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
APC Rat IgG2a, κ Isotype Control RUO
Size
50 Tests
Cat No.
551442
APC Rat Anti-Human IL-2 RUO
Size
25 µg
Cat No.
561054
FITC Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
555916
551383 Rev. 3
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.
551383 Rev. 3
Format Details
APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 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.
APC
Red 627-640 nm
651 nm
660 nm
551383 Rev.3
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. (Clone-specific). 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
551383 Rev. 3
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.
