PDF《cytokines.》

96 well plate. Following this preincubation period, mouse L929 cells are added. ?2012 Sigma-Aldrich Co. LLC. All rights reserved. SIGMA-ALDRICH is a trademark of Sigma-Aldrich Co. LLC, registered in the US and other countries. Sigma brand products are sold through Sigma-Aldrich, Inc. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see product information on the Sigma-Aldrich website at www.sigmaaldrich.com and/or on the reverse side of the invoice or packing slip. The assay mixture is then incubated in a humidified CO2 incubator. The exact concentration of antibody required to neutralize recombinant human TNF-α activity is dependent on the cytokine concentration, cell type, growth conditions, and the type of activity. The Neutralization Dose50 (ND50) for this antibody is defined as that concentration required to yield one-half maximal inhibition of the TNF-α activity on a responsive cell line, when TNF-α is present at a concentration just high enough to elicit a maximum response. Product Profile Capture ELISA: a working antibody concentration of

4 ?g/mL is recommended to detect recombinant human TNF-α. Neutralization: the antibody neutralizes the biological activity of recombinant human TNF-α in a mouse fibrosarcoma cell line (L929). Immunoblotting: a working antibody concentration of 1-2 ?g/mL detects human TNF-α at ~5 ng/lane under non-reducing and reducing conditions. Immunohistochemistry: a minimum working antibody concentration of

25 ?g/mL is recommended for detecting human TNF-α in fixed human peripheral blood leukocytes using the appropriate secondary reagents and chromogenic detection system. Note: In order to obtain the best results in various techniques and preparations, we recommend determining the optimal working dilutions by titration. Endotoxin: < 0.1 EU (endotoxin units) per

1 ?g of the antibody as determined by the LAL method. References 1. Ware, C., et al., Tumor necrosis factor-related ligands and receptors, in The Cytokine Handbook,

3 rd Edition, Thomson, A.W., ed., Academic Press (San Diego, CA: 1998), pp. 549-592. 2. Aggarwal, B., and Reddy, S., Tumor necrosis factor (TNF), in Guidebook to Cytokines and Their Receptors, Nicola, N., ed., Oxford Press (New York, NY: 1994), pp. 103-104. 3. Callard, R., and Gearing, A., The Cytokine Facts Book, Academic Press (New York, NY: 1994). 4. Beutler, B., Cachectin/tumor necrosis factor and lymphotoxin, in Peptide Growth Factors and their Receptors II, Sporn, M., and Roberts, A., eds., Springer-Verlag, (New York, NY: 1991), pp. 39-70. 5. Beutler, B., and Cerami, A., The history, properties, and biological effects of cachectin. Biochemistry, 27, 7575-7582 (1988). 6. Matthews, N., et al., 1987, Lymphokines and Interferons, A Practical Approach, Clemens, A.G., et al., eds., IRL Press, p. 221. TD,PHC 09/12-1