Recombinant Human 4-1BB Ligand/TNFSF9 Protein, CF

• Reactivity: Hu
• Applications: Binding Activity
• Format: Carrier-Free

Recombinant Human 4-1BB Ligand/TNFSF9 Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human (rh) 4-1BB/TNFRSF9/CD137 Fc Chimera (Catalog # 838-4B) is Immobilized at 10 ng/mL (100 µL/well), the concentration of rh4-1BB Ligand that produces 50% optimal binding response is found to be approximately 0.5‑2.5 ng/mL.
• Source: E. coli-derived human 4-1BB Ligand/TNFSF9 protein
  Arg71-Glu254, with an N-terminal Met and a 6-His tag
• Accession #: P41273.1
• N-terminal Sequence: Met
• Protein/Peptide Type: Recombinant Proteins
• Gene: TNFSF9
• Purity: >97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Binding Activity
• Theoretical MW: 20.4 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, 2 to 8 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS, NaCl and PEG and with Trehalose.
• Purity: >97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human 4-1BB Ligand/TNFSF9 Protein, CF

• 41BB Ligand
• 4-1BB Ligand
• 4-1BBL
• 4-1BB-L
• CD137L
• homolog of mouse 4-1BB-L
• receptor 4-1BB ligand
• TNFSF9
• tumor necrosis factor (ligand) superfamily, member 9
• tumor necrosis factor ligand superfamily member 9

Background

4-1BB Ligand (4-1BBL), also known as CD137L, is a type II transmembrane protein that belongs to the TNF superfamily of molecules and plays an important role in immune response activation (1). The 38 kDa human 4-1BB Ligand consists of an 25 aa cytoplasmic domain, a 23 aa transmembrane segment, and a 206 aa extracellular domain (ECD) (2). Within the ECD, human 4-1BB Ligand shares 34% aa sequence identity with mouse and rat 4-1BB Ligand. 4-1BB Ligand is expressed by activated B cells, monocytes, macrophages, dendritic cells (DC), T cells, lymphoma and multiple myeloma cells, hematopoietic stem cells, early myeloid progenitors, neurons, and astrocytes (3-9). A 26 kDa soluble form of 4-1BB Ligand can be released from the surface of activated cells and retains bioactivity (10).
4-1BB Ligand binds to 4-1BB/TNFRSF9/CD137 on activated CD4⁺ and CD8⁺ T cells, thymocytes, and NK cells as well as on monocytes, neutrophils, DC, and eosinophils. In response to 4-1BB Ligand binding, 4-1BB transduces a co-stimulatory signal that promotes the proliferation, activation, and survival of CD4⁺ and CD8⁺ T cells (4, 11, 12). T cell co-stimulation through CD28 is important for the initial T cell expansion, while 4-1BB acts later in the response (12, 13). 4-1BB Ligand function supports the survival and responsiveness of memory T cells during viral infection (13-15). Reverse signaling through 4-1BB Ligand on monocytes induces the production of inflammatory cytokines (5). On macrophages, 4-1BB Ligand associates in cis with TLR4 and enhances inflammatory cytokine production in response to TLR4 ligation (6). Its expression on early myeloid progenitor cells limits the development of dendritic cells, monocytes, and B cells (9).

1. Wang, C. et al. (2009) Immunol. Rev. 229:192.
2. Alderson, M.R. et al. (1994) Eur. J. Immunol. 24:2219.
3. Reali, C. et al. (2003) J. Neurosci. Res. 74:67.
4. DeBenedette, M.A. et al. (1997) J. Immunol. 158:551.
5. Langstein, J. et al. (1998) J. Immunol. 160:2488.
6. Kang, Y.J. et al. (2007) Nat. Immunol. 8:601.
7. Baessler, T. et al. (2010) Blood 115:3058.
8. Gullo, C. et al. (2010) PLoS One 5:e10845.
9. Lee, S.-W. et al. (2008) Nat. Immunol. 9:917.
10. Salih, H.R. et al. (2001) J. Immunol. 167:4059.
11. Wen, T. et al. (2002) J. Immunol. 168:4897.
12. Cannons, J.L. et al. (2001) J. Immunol. 167:1313.
13. Bertram, E.M. et al. (2002) J. Immunol. 168:3777.
14. Bukczynski, J. et al. (2004) Proc. Natl. Acad. Sci. USA 101:1291.
15. Lin, G.H.Y. et al. (2009) J. Immunol. 182:934.

Publications for 4-1BB Ligand/TNFSF9 (2295-4L/CF)(5)

Publications using 2295-4L/CF

• SM Chin, CR Kimberlin, Z Roe-Zurz, P Zhang, A Xu, S Liao-Chan, D Sen, AR Nager, NS Oakdale, C Brown, F Wang, Y Yang, K Lindquist, YA Yeung, S Salek-Arda, J Chaparro-R Structure of the 4-1BB/4-1BBL complex and distinct binding and functional properties of utomilumab and urelumab Nat Commun, 2018-11-08;9(1):4679. 2018-11-08 [PMID: 30410017] (Surface Plasmon Resonance, Human)
• AW Clarke, L Poulton, D Shim, D Mabon, D Butt, M Pollard, V Pande, J Husten, J Lyons, C Tian, AG Doyle An anti-TL1A antibody for the treatment of asthma and inflammatory bowel disease MAbs, 2018-03-05;0(0):1-43. 2018-03-05 [PMID: 29436901] (Surface Plasmon Resonance (SPR, Human)
• Madireddi S, Eun S, Lee S, Nemcovicova I, Mehta A, Zajonc D, Nishi N, Niki T, Hirashima M, Croft M Galectin-9 controls the therapeutic activity of 4-1BB-targeting antibodies. J Exp Med, 2014-06-23;211(7):1433-48. 2014-06-23 [PMID: 24958847] (Bioassay, Human)
• Moh M, Lorenzini P, Gullo C, Schwarz H Tumor necrosis factor receptor 1 associates with CD137 ligand and mediates its reverse signaling. FASEB J, 2013-04-25;27(8):2957-66. 2013-04-25 [PMID: 23620528] (Bioassay, Human)
• Lobito AA, Ramani SR, Tom I, Bazan JF, Luis E, Fairbrother WJ, Ouyang W, Gonzalez LC Murine insulin growth factor-like (IGFL) and human IGFL1 proteins are induced in inflammatory skin conditions and bind to a novel tumor necrosis factor receptor family member, IGFLR1. J. Biol. Chem., 2011-03-31;286(21):18969-81. 2011-03-31 [PMID: 21454693] (Surface Plasmon Resonance, Human)

Bioinformatics

• Gene Symbol: TNFSF9
• Uniprot:
  • P41273.1()