Clone |
MAb11 |
Isotype |
Mouse IgG1, κ |
Reactivity |
Human, Cross-Reactivity: Chimpanzee, Baboon, Cynomolgus, Rhesus, Pigtailed Macaque, Sooty Mangabey, Swine (Pig, Porcine) |
Immunogen |
E. coli-expressed, recombinant human TNF-α |
Formulation |
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide, 0.2% (w/v) BSA (USA origin). |
Preparation |
The antibody was purified by affinity chromatography, and conjugated with Alexa Fluor® 647 under optimal conditions. The solution is free of unconjugated Alexa Fluor® 647. |
Recommended Usage |
Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is 5 µl per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm. ** Alexa Fluor® is a registered trademark of Molecular Probes, Inc. Alexa Fluor® 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. |
Applications |
ICFC-Quality tested |
Application Notes |
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. 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 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 " |
Description |
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. |
Storage |
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. |