Expression of IL-13 by stimulated human lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 6 h with PMA (Sigma, Cat. No. P-8139) and calcium ionophore A23187 (Sigma, Cat. No. C-9275), in the presence of GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). The cells were fixed, permeabilized, and then stained with Alexa Fluor® 488 Mouse anti-Human CD4 antibody (Cat No. 557695) and APC Rat anti-Human IL-13 antibody (Cat No. 561162, Left Panel) or APC Rat IgG1 Isotype Control (Cat. No. 554686, Right Panel) by using BD Biosciences' protocol, Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color flow cytometric dot plots were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
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Expression of IL-13 by stimulated human lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 6 h with PMA (Sigma, Cat. No. P-8139) and calcium ionophore A23187 (Sigma, Cat. No. C-9275), in the presence of GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). The cells were fixed, permeabilized, and then stained with Alexa Fluor® 488 Mouse anti-Human CD4 antibody (Cat No. 557695) and APC Rat anti-Human IL-13 antibody (Cat No. 561162, Left Panel) or APC Rat IgG1 Isotype Control (Cat. No. 554686, Right Panel) by using BD Biosciences' protocol, Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color flow cytometric dot plots were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
Interleukin-13; NC30; ALRH; BHR1; P600
Human (QC Testing)
Rat IgG1
Human Recombinant IL-13
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
3596
AB_10642586
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Companion Products
Fixation/Permeabilization Kit RUO
Size
250 Tests
Cat No.
554714
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 mL
Cat No.
554724
APC Rat IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554686
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Alexa Fluor®488 Mouse Anti-Human CD4 RUO
Size
100 Tests
Cat No.
557695
561162 Rev. 1
Antibody Details
JES10-5A2
The JES10-5A2 monoclonal antibody specifically binds to human interleukin-13, IL-13. IL-13 is produced by activated T cells, mast cells and NK cells. IL-13 regulates IgE production by B cells and can suppress the cytotoxic activity of macrophages and their production of inflammatory mediators. The immunogen used to produce the JES10-5A2 hybridoma was COS-expressed recombinant human IL-13. This is a neutralizing antibody.
561162 Rev. 1
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
561162 Rev.1
Citations & References
Development References (3)
-
Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Clone-specific: ELISA). View Reference
-
McKenzie A, Zurawski G. Measurement of IL-13. In: Coligan, Kruisbeek, Shevak, Strober, ed. Current Protocols in Immunology. New York: John Wiley & Sons; 1994:18-19.
-
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: IC/FCM Block). View Reference
561162 Rev. 1
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.
