BD Pharmingen™ Biotin Human anti-Mouse/Rat TNF
Clone 516D1A1 (RUO)
Mouse TNF sandwich ELISA curve: The curve was generated by sandwich ELISA using the purified TN3-19.12 (Cat. No. 557516) as capture antibody, doubling dilutions of recombinant mouse TNF protein (Cat. No. 554589) and biotin 516D1A1 as detection antibody. Streptavidin-HRP (Cat. No. 554066) plus TMB substrate (Cat. No. 555214) were used to develop the ELISA.
Mouse TNF sandwich ELISA curve: The curve was generated by sandwich ELISA using the purified TN3-19.12 (Cat. No. 557516) as capture antibody, doubling dilutions of recombinant mouse TNF protein (Cat. No. 554589) and biotin 516D1A1 as detection antibody. Streptavidin-HRP (Cat. No. 554066) plus TMB substrate (Cat. No. 555214) were used to develop the ELISA.
Preparation And Storage
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Companion Products
The 516D1A1 Fab antibody reacts with mouse and rat tumor-necrosis factor (TNF, formerly known as TNF-α) proteins. The 516D1A1 Fab was selected by screening a human Fab phage display library (Hoet et al.) licensed from Dyax Corp (Cambridge, MA) with recombinant mouse TNF protein. The biology and function of the TNF protein has been extensively reviewed (Hehlgans et al.) in the literature.
The biotinylated 516D1A1 antibody is useful as a detection antibody for a sandwich ELISA that measures mouse and rat TNF protein levels. Purified Hamster Anti-Mouse/Rat TNF (Cat. No.557516) as capture antibody can be paired with biotinylated 516D1A1 as the detection antibody, using purified recombinant mouse TNF protein (Cat. No. 554589) or purified recombinant rat TNF protein (Cat. No. 555109) as standard. The biotinylated 516D1A1 antibody should be titrated between 0.25 - 2.0 µg/ml to determine its optimal ELISA detection concentration. To obtain linear standard curves, doubling dilutions of purified mouse or rat TNF protein ranging from 31.25 pg/ml to 2,000 pg/ml are recommended for inclusion in each ELISA plate. Investigators may find methodology information to be useful in Techniques for Immune Function Analysis Application Handbook 1st Edition. BD Biosciences (2003)-- Chapter 7: ELISA for specifically measuring the levels of cytokines, chemokines, and inflammatory mediators and their receptors. For this ELISA antibody pair, no cross-reactivity has been observed with the following: Rat IL-1α, IL-2, IL-4, IL-6, IL-10, IL-18, GM-CSF, IFN-γ; mouse IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-18, M-CSF, GM-CSF, IFN-γ, MCP-1, MIP-1α, MIP-1β, sTNFRI, sTNFRII, RANTES; human TNF.
Development References (2)
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Hehlgans T, Pfeffer K. The intriguing biology of the tumour necrosis factor/tumour necrosisfactor receptor superfamily: players, rules and the games. Immunology. 2005; 115(1):1-20. (Biology). View Reference
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Hoet RM, Cohen EH, Kent RB, et al. Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity. Nat Biotechnol. 2005; 23(3):344-348. (Clone-specific: ELISA). View Reference
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