Recombinant Mouse Lymphotoxin α1/β2 Catalog Number: 9968-LY/CF DESCRIPTION Source Mouse myeloma cell line, NS0­derived mouse Lymphotoxin protein Mouse LT alpha Mouse LT beta Mouse LT beta (Lys59­Leu202) GGGGS (Leu153­Gly306) GGGGS (Leu153­Gly306) Accession # P09225 Accession # P41155 Accession # P41155 N­terminal Sequence Lys59 Analysis Structure / Form GS­linked heterotrimer Predicted Molecular 50 kDa Mass SPECIFICATIONS SDS­PAGE 42­63 kDa, reducing conditions Activity Measured in a cell proliferation assay using NIH­3T3 mouse embryonic fibroblast cells. The ED50 for this effect is 0.3­2.1 ng/mL Endotoxin Level <0.10 EU per 1 μg of the protein by the LAL method. Purity >95%, by SDS­PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. Formulation Lyophilized from a 0.2 μm filtered solution in PBS. See Certificate of Analysis for details. PREPARATION AND STORAGE Reconstitution Reconstitute at 500 μg/mL in PBS. Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. Stability & Storage l 12 months from date of receipt, ≤ ­20 °C as supplied. l 1 month, 2 to 8 °C under sterile conditions after reconstitution. l 3 months, ≤ ­20 °C under sterile conditions after reconstitution. DATA Bioactivity SDS­PAGE Recombinant Mouse 2 μg/lane of Recombinant Mouse Lymphotoxin alpha1/beta2 Lymphotoxin α1/β2 was resolved with SDS­PAGE under reducing (R) and non­ (Catalog # 9968­ reducing (NR) conditions and visualized by Coomassie® blue LY/CF) induces NIH­ staining, showing bands at 42­63 kDa. 3T3 mouse embryonic fibroblast cell proliferation. The ED50 for this effect is 0.3­2.1 ng/mL. Rev. 6/26/2018 Page 1 of 2 Recombinant Mouse Lymphotoxin α1/β2 Catalog Number: 9968-LY/CF BACKGROUND Lymphotoxin alpha (LT alpha) and Lymphotoxin beta (LT beta) are pro­inflammatory TNF superfamily ligands that play important roles in immune system development (1, 2). Mouse LT alpha cDNA encodes a 202 amino acid (aa) secreted soluble protein with a 33 aa signal sequence. The mature mouse LT alpha shares 75% aa sequence identity with human LT alpha (3, 4). Mouse LT beta cDNA encodes a 306 aa type II membrane protein with a 27 aa N­terminal cytoplasmic domain, a 21 aa transmembrane region, and a 258 aa extracellular domain. It shares 73% aa sequence identity with human LT beta within common regions of their extracellular domains (5). Secreted LT alpha assembles as a soluble homotrimer, LT alpha 3. In addition, secreted LT alpha also complexes with the membrane associated LT beta to generate two types of heterotrimers, LT alpha 1/beta 2 and LT alpha 2/beta 1 (5). The soluble LT alpha 3 binds both TNF RI (p55) and TNF RII (p75). In contrast, the predominant membrane­bound heterotrimer, LT alpha 1/beta 2, binds only to the lymphotoxin beta receptor (LT beta R). LT alpha 2/beta 1 is capable of binding LT beta R, TNF RI (p55), and TNF RII (p75). LT plays a role in normal lymphoid organogenesis (6, 7). The LT alpha 1/beta 2 heterotrimer binds and activates the LT beta R/TNFRSF3 (LT beta R) which is expressed on macrophages, dendritic cells, hepatocytes, intestinal epithelial cells (IEC), follicular dendritic cells (FDC), and high endothelial venules (HEV) (2, 8, 9). LT beta R also serves as a receptor for LIGHT/TNFSF14 (10). LT alpha 1/beta 2 promotes the development of FDC networks and HEV in lymphoid tissue, the class switching of immature B cells for IgA production, and the production of homeostatic IL­22 by ILCs (11­14). It can be shed by ADAM17 or MMP­8 mediated cleavage, and the released heterotrimer circulates in the serum and is elevated in synovial fluid of rheumatoid arthritis patients (15). References: 1. Lu, T.T. and J.L. Browning (2014) Front. Immunol. 5:47. 2. Upadhyay, V. and Y.­X. Fu (2014) Cytokine Growth Factor Rev. 25:227. 3. Gray, P.W. et al. (1984) Nature 312:721. 4. Nedwin, G.E. et al. (1985) J. Cell Biochem. 29:171. 5. Browning, J.L. et al. (1993) Cell 72:847. 6. Ettinger R. et al. (1996) Proc. Natl. Acad. Sci. USA 93:13102. 7. Cuff, C.A. et al. (1998) J. Immunol. 161:6853. 8. Sudhamsu, J. et al. (2013) Proc. Natl. Acad. Sci. USA 110:19896. 9. Crowe, P.D. et al. (1994) Science 264:707. 10. Eldredge, J. et al. (2006) Biochemistry 45:10117. 11. Kruglov, A.A. et al. (2013) Science 342:1243. 12. Futterer, A. et al. (1998) Immunity 9:59. 13. Koni, P.A. et al. (1997) Immunity 6:491. 14. Ota, N. et al. (2011) Nat. Immunol. 12:941. 15. Young, J. et al. (2010) Cytokine 51:78. Rev. 6/26/2018 Page 2 of 2 .