The JES6-5H4 antibody reacts with mouse interleukin-2 (IL-2) and has been reported to be neutralizing. Expression of IL-2 by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from 6 month old BALB/c mice were stimulated for 5 hours with hamster anti-mouse CD3 (2 μg/mL final concentration, clone 145-2C11, Cat. No. 553057) and hamster anti-mouse CD28 (2 μg/mL final concentration, clone 37.51, Cat. No. 553294) antibodies in the presence of BD GolgiStop™ (3 μM final concentration, Cat. No. 554724). The cells were harvested, stained with 0.06 μg of PE-conjugated rat anti-mouse CD4 (PE-RM4-5, Cat. No. 553049), fixed, permeabilized, and subsequently stained with 0.06 μg of FITC-conjugated rat anti-mouse IL-2 antibody (FITC-JES6-5H4, Cat. No. 554427, left panel). To demonstrate specificity of staining, the binding by the FITC-JES6-5H4 antibody was blocked by preincubation of the conjugated antibody with recombinant mouse IL-2 (0.12 μg, Cat. No. 550069, middle panel), and by preincubation of the fixed/permeabilized cells with unlabeled JES6-5H4 antibody (2.5 μg, Cat. No. 554425, right panel) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabelled antibody blocking specificity controls.
BD Pharmingen™ Purified Rat Anti-Mouse IL-2 - Purified - Clone JES6-5H4 - Isotype Rat IgG2b - Reactivity Ms - 0.1 mg
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C
Storage Comment
Store undiluted at 4°C.
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Prussin, Metcalfe: "Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies." in: Journal of immunological methods, Vol. 188, Issue 1, pp. 117-28, (1996) (PubMed).
Abrams, Roncarolo, Yssel, Andersson, Gleich, Silver: "Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples." in: Immunological reviews, Vol. 127, Issue 9-10, pp. 5-24, (1992) (PubMed).