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The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤ 0.5 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome. ** Pacific Blue™ is a registered trademark of Molecular Probes, Inc. Pacific Blue™ dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
ELISA or ELISPOT Detection: The biotinylated MAb11 antibody is useful as the detection antibody in a sandwich ELISA or ELISPOT, when used in conjunction with the purified MAb1 antibody (Cat. No. 502802/502804) as the capture antibody. Flow Cytometry3,5,6: The fluorochrome-labeled MAb11 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify TNF-α -producing cells within mixed cell populations. View intracellular cytokine staining protocol Additional reported applications (for the relevant formats) include: neutralization1,2, immunohistochemical staining of paraformaldehyde-fixed, saponin-treated frozen tissue sections4, and immunocytochemistry. The MAb11 antibody can neutralize the bioactivity of natural or recombinant TNF-α. Note: For testing human TNF-α in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 430201 to 430206) are specially developed and recommended. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for neutralization of human TNF-α bioactivity (Cat. No. 502922).
Application References:
1. Rathjen D, et al. 1991. Mol. Immunol. 28:79. (Neut) 2. Danis V, et al. 1991. Clin. Exp. Immunol. 85:143. (Neut) 3. Enr quez J, et al. 2002. Adv. Perit. Dial. 18:177. (ICFC) 4. Andersson U, et al. 1999. Detection and quantification of gene expression. New York:Springer-Verlag. (IHC) 5. Chen H, et al. 2005. J. Immunol. 175:591. (ICFC) 6. Iwamoto S, et al. 2007. J. Immunol. 179:1449. (ICFC) PubMed
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with mouse IgG1 Pacific Blue™ isotype control and CD3 (UCHT1) APC
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with MAb11 Pacific Blue™ and CD3 (UCHT1) APC
TNF-α is secreted by macrophages, monocytes, neutrophils, T-cells (principally CD4+), and NK-cells. Many transformed cell lines also secrete TNF-α. Monomeric human TNF-α is a 157 amino acid protein (non-glycosylated) with a reported molecular weight of 17 kD. TNF-α forms multimeric complexes; stable trimers are most common in solution. A 26 kD membrane form of TNF-α has also been described. TNF-α binding to surface receptors elicits a wide array of biologic activities including: cytolysis and cytostasis of many tumor cell lines in vitro, hemorraghic necrosis of tumors in vivo, increased fibroblast proliferation, and enhanced chemotaxis and phagocytosis in neutrophils.
Type II integral membrane protein processed by TACE for secretion; upregulated by interferons, IL-2, GM-CSF, substance P, bradykinin, PAF, immune complexes, cyclooxygenase; downregulated by IL-6, TGF-β, vitamin D3, prostaglandin E2, PAF antagonists
Cellular Sources:
Activated monocytes, neutrophils, macrophages, T cells, B cells, NK cells, LAK cells
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
APC anti-human TNF-α
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with mouse IgG1 APC isotype control and CD3 (UCHT1) PE
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with MAb11 APC and CD3 (UCHT1) PE
Biotin anti-human TNF-α
FITC anti-human TNF-α
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with MAb11 FITC and CD3 (UCHT1) APC
PE anti-human TNF-α
PMA/Ionomycin-stimulated human
PBMCs were stained with CD3
PE/Cy5 and MAb11 PE
Alexa Fluor® 488 anti-human TNF-α
PMA+ionomycin-stimulated (6 hours) human peripheral blood lymphocytes intracellular stained with MAB11 Alexa Fluor®
488 and CD3 (UCHT1) PE
Alexa Fluor® 647 anti-human TNF-α
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with mouse IgG1 Alexa Fluor® 647 isotype control and CD3 (UCHT1) PE
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with MAb11 Alexa Fluor® 647 and CD3 (UCHT1) PE
Pacific Blue™ anti-human TNF-α
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with mouse IgG1 Pacific Blue™ isotype control and CD3 (UCHT1) APC
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with MAb11 Pacific Blue™ and CD3 (UCHT1) APC
PerCP/Cy5.5 anti-human TNF-α
PMA + ionomycin-stimulated (6 hours) human peripheral blood lymphocytes intracellular stained with MAB11 PerCP/Cy5.5 and CD3 (UCHT1) PE
PE/Cy7 anti-human TNF-α
PMA+ionomycin-stimulated (6 hours) peripheral blood lymphocytes surface stained with CD3 FITC, then intracellularly stained with MAB11 PE/Cy7 (top) or mouse IgG1,k PE/Cy7 isotype control (bottom)
Brilliant Violet 421™ anti-human TNF-α
PMA+ionomycin stimulated (6 hours) human peripheral blood lymphocytes (in the presence of monensin) were stained with CD3 FITC, fixed, permeabilized, and then stained with TNF-α (clone MAb11) Brilliant Violet 421™ (top) or mouse IgG1, κ Brilliant Violet 421™ isotype control (bottom).
