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BD Pharmingen™ PE Mouse Anti-Human TNF
Clone MAb11 (RUO)
Expression of TNF protein by stimulated human peripheral blood mononuclear cells (PBMC). Human 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) and PHA (Gibco BRL, Cat. #10576-015) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554715). The PBMC were stained with 0.25 µg PE-Cy™5 Mouse Anti-Human CD3 (Cat. No. 555334) and subsequently stained with 0.5 µg of PE Mouse Anti-Human TNF (Cat. No. 554513/557068/559321/562083; left panel). To demonstrate specificity of staining, the binding of PE Mouse Anti-Human TNF was blocked by preincubation of the conjugate with excess (1 µg) recombinant human TNF (Cat. No. 554618; middle panel) and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF (Cat. No. 554510; right panel) prior to staining with the PE Mouse Anti-Human TNF. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls.
Expression of TNF protein by stimulated human peripheral blood mononuclear cells (PBMC). Human 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) and PHA (Gibco BRL, Cat. #10576-015) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554715). The PBMC were stained with 0.25 µg PE-Cy™5 Mouse Anti-Human CD3 (Cat. No. 555334) and subsequently stained with 0.5 µg of PE Mouse Anti-Human TNF (Cat. No. 554513/557068/559321/562083; left panel). To demonstrate specificity of staining, the binding of PE Mouse Anti-Human TNF was blocked by preincubation of the conjugate with excess (1 µg) recombinant human TNF (Cat. No. 554618; middle panel) and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF (Cat. No. 554510; right panel) prior to staining with the PE Mouse Anti-Human TNF. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls.
Expression of TNF protein by stimulated human peripheral blood mononuclear cells (PBMC). Human 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) and PHA (Gibco BRL, Cat. #10576-015) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554715). The PBMC were stained with 0.25 µg PE-Cy™5 Mouse Anti-Human CD3 (Cat. No. 555334) and subsequently stained with 0.5 µg of PE Mouse Anti-Human TNF (Cat. No. 554513/557068/559321/562083; left panel). To demonstrate specificity of staining, the binding of PE Mouse Anti-Human TNF was blocked by preincubation of the conjugate with excess (1 µg) recombinant human TNF (Cat. No. 554618; middle panel) and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF (Cat. No. 554510; right panel) prior to staining with the PE Mouse Anti-Human TNF. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls.
Expression of TNF protein by stimulated human peripheral blood mononuclear cells (PBMC). Human 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) and PHA (Gibco BRL, Cat. #10576-015) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554715). The PBMC were stained with 0.25 µg PE-Cy™5 Mouse Anti-Human CD3 (Cat. No. 555334) and subsequently stained with 0.5 µg of PE Mouse Anti-Human TNF (Cat. No. 554513/557068/559321/562083; left panel). To demonstrate specificity of staining, the binding of PE Mouse Anti-Human TNF was blocked by preincubation of the conjugate with excess (1 µg) recombinant human TNF (Cat. No. 554618; middle panel) and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF (Cat. No. 554510; right panel) prior to staining with the PE Mouse Anti-Human TNF. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls.
제품 세부 정보
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 staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_395444
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
준비 및 보관
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
권장 분석 절차
Immunofluorescent Staining and Flow Cytometric Analysis: The MAb11 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify and enumerate TNF producing cells within mixed cell populations (see image). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be pretitrated (≤ 1 µg mAb/million cells). The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit the protocols section under "Cytokines (Intracellular Staining)" or "Intracellular Flow" at our website, http://www.bdbiosciences.com/us/s/resources.
제품 고시
- 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.
- 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.
관련 제품
Stain Buffer (FBS) RUO
사이즈
500 ML
카탈로그 No
554656
Stain Buffer (BSA) RUO
사이즈
500 ML
카탈로그 No
554657
PE Mouse IgG1, κ Isotype Control RUO
사이즈
0.1 mg
카탈로그 No
554680
Recombinant Human TNF RUO
사이즈
10 µg
카탈로그 No
554618
PE Mouse Anti-Human TNF RUO
사이즈
100 Tests
카탈로그 No
559321
PE Mouse Anti-Human TNF RUO
사이즈
25 Tests
카탈로그 No
562083
554513 Rev. 3
항체 세부 정보
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.
554513 Rev. 3
형광 세부 정보
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
496 nm, 566 nm
576 nm
554513 Rev.3
인용 및 참고 문헌
View product citations for antibody "554513" on CiteAb
개발 참고 자료 (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). 참조 보기
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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). 참조 보기
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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: Flow cytometry). 참조 보기
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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). 참조 보기
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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). 참조 보기
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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). 참조 보기
554513 Rev. 3
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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.