Expression of IL-4 by stimulated Rhesus macaque peripheral blood lymphocytes. Rhesus macaque peripheral blood mononuclear cells were stimulated for 6 h with Phorbol 12-Myristate 13-Acetate (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 using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with FITC Mouse Anti-Human CD3 (Cat No. 556611) and APC Mouse anti-Human IL-4 antibody (Cat No. 561233, Left Panel), or APC Mouse IgG1 κ Isotype Control (Cat No.554681, Right Panel) by using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
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Expression of IL-4 by stimulated Rhesus macaque peripheral blood lymphocytes. Rhesus macaque peripheral blood mononuclear cells were stimulated for 6 h with Phorbol 12-Myristate 13-Acetate (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 using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with FITC Mouse Anti-Human CD3 (Cat No. 556611) and APC Mouse anti-Human IL-4 antibody (Cat No. 561233, Left Panel), or APC Mouse IgG1 κ Isotype Control (Cat No.554681, Right Panel) by using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
Interleukin-4; IL4; BCGF-1; BSF-1; Lymphocyte stimulatory factor 1
Rhesus, Cynomolgus (QC Testing)
Mouse IgG1, κ
Recombinant Human IL-4
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_10611724
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.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 mL
Cat No.
554724
APC Mouse IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
554681
FITC Mouse Anti-Human CD3ε RUO
Size
50 Tests
Cat No.
556611
561233 Rev. 2
Antibody Details
8D4-8
The 8D4-8 monoclonal antibody reacts with human interleukin-4 (IL-4). The immunogen used to raise the 8D4-8 hybridoma was recombinant human IL-4. The 8D4-8 antibody binds to an epitope that is different than the epitope recognized by the MP4-25D2 antibody (Cat. No. 554485).
Clone 8D4-8 displays an increased amount of non-specific binding to dead cells when compared to the clone MP4-25D2. It is recommended to use a fixable viability dye in conjunction with this clone.
561233 Rev. 2
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
561233 Rev.2
Citations & References
Development References (2)
-
Callard R, Gearing A. Callard R, Gearing A. The Cytokine Facts Book. San Diego: Academic Press; 1994.
-
Paul WE. Interleukin-4: a prototypic immunoregulatory lymphokine. Blood. 1991; 77(9):1859-1870. (Biology). View Reference
561233 Rev. 2
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.
