Expression of TNF protein by stimulated Rhesus macaque peripheral blood mononuclear cells (PBMC). Rhesus macaque PBMC were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. #P-8139), calcium ionophore A23187 (0.5 μg/ml final concentration; Sigma, Cat. #C-9275) in the presence of GolgiStop™ (2 μM final concentration; Cat. No. 554715). The PBMC were stained with either Purified Mouse Anti-Human TNF (Cat. No. 558882/554510; solid line histogram) or Purified Mouse IgG1 κ Isotype Control (Cat. No. 556648; dashed line histogram). Secondary staining was carried out with FITC Goat Anti-Mouse IgG/IgM (Cat. No. 555988).Fluorescence histograms were derived from gated events with the side and forward light-scatter characteristics of viable lymphocytes.
Expression of TNF protein by stimulated Rhesus macaque peripheral blood mononuclear cells (PBMC). Rhesus macaque PBMC were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. #P-8139), calcium ionophore A23187 (0.5 μg/ml final concentration; Sigma, Cat. #C-9275) in the presence of GolgiStop™ (2 μM final concentration; Cat. No. 554715). The PBMC were stained with either Purified Mouse Anti-Human TNF (Cat. No. 558882/554510; solid line histogram) or Purified Mouse IgG1 κ Isotype Control (Cat. No. 556648; dashed line histogram). Secondary staining was carried out with FITC Goat Anti-Mouse IgG/IgM (Cat. No. 555988).Fluorescence histograms were derived from gated events with the side and forward light-scatter characteristics of viable lymphocytes.
Product Details
BD Pharmingen™
Tumor necrosis factor alpha; TNF-a; TNF-α; TNFSF2; Cachectin
Human (QC Testing), Rhesus,Cynomolgus,Baboon (Tested in Development)
Mouse IgG1, κ
Recombinant Human TNF
Intracellular block/flow cytometry (Routinely Tested)
0.5 mg/ml
AB_397142
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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
Purified Mouse IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
556648
FITC Goat Anti-Mouse IgG/IgM RUO
Size
0.5 mg
Cat No.
555988
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558882 Rev. 9
Antibody Details
MAb11
The MAb11 monoclonal antibody specifically binds to human tumor necrosis factor (TNF, also known as TNF-α) protein. TNF is an efficient juxtacrine, paracrine and endocrine mediator of inflammatory and immune functions. It regulates the growth and differentiation of a variety of cell types. TNF is cytotoxic for transformed cells when in conjunction with IFN-γ. It is secreted by activated monocytes/macrophages and other cells such as B cells, T cells and fibroblasts. The immunogen used to generate the MAb11 hybridoma was recombinant human TNF. The MAb11 antibody has been reported to crossreact with Rhesus Macaque TNF.
558882 Rev. 9
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
558882 Rev.9
Citations & References
Development References (8)
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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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Jason J, Larned J. Single-cell cytokine profiles in normal humans: comparison of flow cytometric reagents and stimulation protocols. J Immunol Methods. 1997; 207(1):13-22. (Biology). 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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Sander B, Andersson J, Andersson U. Assessment of cytokines by immunofluorescence and the paraformaldehyde-saponin procedure. Immunol Rev. 1991; 119:65-93. (Biology). 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
558882 Rev. 9
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.
