Flow cytometric profile of TNF expression on rhesus macaque (Macaca mulatta) peripheral blood lymphocytes. Peripheral blood lymphocytes were stimulated with PMA, then fixed, permeabilized, and washed with the BD Cytofix/Cytoperm Plus Kit (with BD GolgiStop)(Cat. No. 554715). Samples were then costained with APC Mouse Anti-Human TNF (Cat. No. 551384) and FITC Mouse Anti-Human CD3ε (Cat. No. 556611). The two-color dot plot was derived from gated events with the forward and side-light scatter characteristics of viable lymphocytes.
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Flow cytometric profile of TNF expression on rhesus macaque (Macaca mulatta) peripheral blood lymphocytes. Peripheral blood lymphocytes were stimulated with PMA, then fixed, permeabilized, and washed with the BD Cytofix/Cytoperm Plus Kit (with BD GolgiStop)(Cat. No. 554715). Samples were then costained with APC Mouse Anti-Human TNF (Cat. No. 551384) and FITC Mouse Anti-Human CD3ε (Cat. No. 556611). The two-color dot plot was derived from gated events with the forward and side-light scatter characteristics of viable lymphocytes.
Product Details
BD Pharmingen™
Tumor necrosis factor alpha; TNF-a; TNF-α; TNFSF2; Cachectin
Human, Rhesus, Cynomolgus, Baboon (QC Testing)
Mouse IgG1, κ
Recombinant Human TNF
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_2204110
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation/Permeabilization Solution Kit with BD GolgiStop™ RUO
Size
250 Tests
Cat No.
554715
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
551384 Rev. 3
Antibody Details
MAb11
Clone MAb11 reacts with the human form of tumor necrosis factor (TNF, formerly known as TNF-α) detectable in the cytoplasm of a major subset of activated peripheral blood T lymphocytes. Using standard procedures for detection of intracellular proteins, clone MAb11 also cross-reacts with activated peripheral blood CD3+ lymphocytes of baboon, and both rhesus and cynomolgus macaque monkeys following five-hour treatment with phorbol myristic acetate (PMA) and Ca++ Ionophore (A23187) in the presence of monensin. The staining is restricted to CD3+ T cells and is similar to that observed with peripheral blood T lymphocytes from normal human donors.
551384 Rev. 3
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
551384 Rev.3
Citations & References
Development References (6)
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Danis VA, Franic GM, Rathjen DA, Brooks PM. Effects of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-2, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and IL-6 on the production of immunoreactive IL-1 and TNF-alpha by human monocytes. Clin Exp Immunol. 1991; 85(1):143-150. (Clone-specific: ELISA). View Reference
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Meager A. Characterization of interferons and immunoassays. In: Clemens MJ, Morris AG, Gearing AJH, ed. Lymphokines and Interferons. A Practical Approach. Oxford: IRL Press Ltd; 1987:105-127.
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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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Rathjen DA, Cowan K, Furphy LJ, Aston R. Antigenic structure of human tumour necrosis factor: recognition of distinct regions of TNF alpha by different tumour cell receptors. Mol Immunol. 1991; 28(1-2):79-86. (Clone-specific: ELISA). View Reference
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Sopper S, Stahl-Hennig C, Demuth M, Johnston IC, Dorries R, ter Meulen V. Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys. Cytometry. 1997; 29(4):351-362. (Biology). View Reference
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Verdier F, Aujoulat M, Condevaux F, Descotes J. Determination of lymphocyte subsets and cytokine levels in cynomolgus monkeys. Toxicology. 1995; 105(1):81-90. (Biology). View Reference
551384 Rev. 3
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.
