TNF-a, the prototypical member of the TNF protein superfamily, is a homotrimeric type-II membrane protein (1,2). Membrane-bound TNF-a is cleaved by the metalloprotease TACE/ADAM17 to generate a soluble homotrimer (2). Both membrane and soluble forms of TNF-a are biologically active. TNF-a is produced by a variety of immune cells including T cells, B cells, NK cells, and macrophages (1). Cellular response to TNF-a is mediated through interaction with receptors TNF-R1 and TNF-R2 and results in activation of pathways that favor both cell survival and apoptosis depending on the cell type and biological context. Activation of kinase pathways (including JNK, ERK (p44/42), p38 MAPK, and NF-kB) promotes the survival of cells, while TNF-a-mediated activation of caspase-8 leads to programmed cell death (1,2). TNF-a plays a key regulatory role in inflammation and host defense against bacterial infection, notably Mycobacterium tuberculosis (3). The role of TNF-a in autoimmunity is underscored by blocking TNF-a action to treat rheumatoid arthritis and Crohns disease (1,2,4). Applications: Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested. Recommended Dilution: Western Blot: 1:1000, incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4C with gentle shaking, overnight. Immunoprecipitation: 1:100 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
