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Expression of IL-10 by MiCK-2 Cytokine Positive Control Cells. MiCK-2 cells (Cat. No. 554653) were permeabilized, and subsequently stained with 0.12 µg of PE-conjugated rat anti-mouse IL-10 antibody (PE-JES5-16E3, Cat. No. 554467; right panel) or 0.12 µg of PE-A95-1 rat IgG2b isotype control immunoglobulin (Cat. No. 556925; left panel) by using the BD Pharmingen staining protocol. The quadrant markers for the bivariate dot plots were set based on autofluorescence control.
BD Pharmingen™ PE Rat IgG2b, κ Isotype Control
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
Immunofluorescent Staining and Flow Cytometric Analysis: The PE-conjugated A95-1 immunoglobulin (Cat. No. 556925) is a suitable rat IgG2b, κ isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse and human cells for flow cytometric analysis (see right panel). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
Companion Products
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.