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Expression of IL-2 by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA (5 ng/ml, Sigma, Cat. No. P-8139) and Ionomycin (500 ng, Sigma, P-8139) in the presence of BD GolgiPlug™ (Cat. No. 555029). Cells were harvested, fixed, permeabilized, and stained with PE-conjugated rat anti-mouse CD4 (PE-RM4-5, Cat. No. 553048) and either rat anti-mouse IL-2 antibody (Alexa Fluor® 488-JES6-5H4, Cat. No. 557725), (left panel) or immunoglobulin isotype control (Alexa Fluor® 488-A95-1, Cat. No. 557726), (right panel) by using Pharmingen's staining protocol. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls.
BD Pharmingen™ Alexa Fluor® 488 Rat IgG2b, κ Isotype Control
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Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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The A95-1 antibody has unknown specificity. Trinitrophenal (TNP), the immunogen, is a hapten that is not expressed on human, mouse, rat, or non-human primate cells. The A95-1 immunoglobulin was selected as an isotype control following screening for low background on a variety of mouse and human tissues.
Development References (1)
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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
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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.