4-1BBL/TNFSF9 Protein, Mouse (HEK293, His)

4-1BBL (4-1BB ligand) is a type II membrane protein of the TNF superfamily, is expressed on antigen-presenting cells. 4-1BB with 4-1BBL can induce T-cell expansion, cytokine induction, differentiation, and upregulation of anti-apoptotic genes as well as protect T cells from activation-induced cell death (AICD)^[1]. 4-1BBL stimulates T cell proliferation and induces effective anti-tumor immune responses^[3]. 4-1BBL is an immunostimulant molecule that interacts with the 4-1BB high-affinity receptor during the antigen presentation, providing costimulatory signals to both CD4+ and CD8+ T cells through the activation of NF-kB, c-Jun, and p38 downstream pathways, triggering pleiotropic effects on the immune system^[4]. 4-1BBL/TNFSF9 Protein, Mouse (HEK293, His) is a recombinant protein with a N-Terminal His label, It consists of 206 amino acids (R104-E309) and is produced in HEK293 cells.

4-1BBL is expressed on a variety of antigen presenting cells (APCs), including activated B cells, dendritic cells, macrophages, and myeloid cells^[1].
The amino acid sequence of human 4-1BBL protein has low homology for mouse and rat 4-1BBL protein.
4-1BBL binds to high-affinity 4-1BB, resulting in the recruitment of intracellular TRAF adaptor molecules (TRAF1 and TRAF2), and then activate of NF-jB and the extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen-associated protein (MAP) kinase signaling cascades. The binding of 4-1BBL to 4-1BB generates strong co-stimulatory signals in T-cells that lead to up-regulation of anti-apoptotic molecules, cytokine secretion, and enhanced effector function^[2].
4-1BBL is a member of the TNF family of proteins. 4-1BBL is an immunostimulant molecule that interacts with the 4-1BB high-affinity receptor during the antigen presentation, providing costimulatory signals to both CD4+ and CD8+ T cells through the activation of NF-kB, c-Jun, and p38 downstream pathways, triggering pleiotropic effects on the immune system^[4]. 4-1BBL significantly induces T cell proliferation and increases the stimulation of both IL-2 and IFN-γ^[5].

4-1BB (hamster ovary; 5 μg/mL; 16 h) costimulates B lymphocyte proliferation^[4].

1.Measured by its ability to co-stimulate IL-2 secretion by mouse T cells in the presence of anti-CD3. The ED₅₀ for this effect is 1-10 ng/mL.
2.Measured by its binding ability in a functional ELISA. Immobilized Recombinant Mouse 4-1BBL at 2µg/mL (100 µL/well) can bind biotinylated Mouse 4-1BB. The ED₅₀ for this effect is 0.4068 μg/mL.

• 
  Measured by its ability to co-stimulate IL-2 secretion by mouse T cells in the presence of anti-CD3.The ED₅₀ for this effect is 2.149 ng/mL, corresponding to a specific activity is 4.65×10⁵ U/mg.

Mouse

HEK293

N-10*His

P41274 (R104-E309)

21950  [NCBI]

Extracellular Domain

HHHHHHHHHHRTEPRPALTITTSPNLGTRENNADQVTPVSHIGCPNTTQQGSPVFAKLLAKNQASLCNTTLNWHSQDGAGSSYLSQGLRYEEDKKELVVDSPGLYYVFLELKLSPTFTNTGHKVQGWVSLVLQAKPQVDDFDNLALTVELFPCSMENKLVDRSWSQLLLLKAGHRLSVGLRAYLHGAQDAYRDWELSYPNTTSFGLFLVKPDNPWE

Approximately 33-45 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

• 
  ≥ 90%, as determined by reducing SDS-PAGE.

Lyophilized powder

1.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.
2.Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 150 mM NaCl, pH 7.4.
3.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 5% trehalose.
4.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.
Please refer to the lot-specific COA for specific buffer information.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.

• [1]. Li Y, et al. Limited Cross-Linking of 4-1BB by 4-1BB Ligand and the Agonist Monoclonal Antibody Utomilumab. Cell Rep. 2018 Oct 23;25(4):909-920.e4.  [Content Brief]

  [2]. Yuan Zhao ,et al. Dispensable Role for 4-1BB and 4-1BBL in Development of Vaccinia Virus-Specific CD8 T Cells. Immunol Lett. 2012 Jan 30;141(2):220-6.  [Content Brief]

  [3]. Bitra A, et al. Crystal structure of the m4-1BB/4-1BBL complex reveals an unusual dimeric ligand that undergoes structural changes upon 4-1BB receptor binding. J Biol Chem. 2019 Feb 8;294(6):1831-1845.  [Content Brief]

  [4]. Meseck M, et al. A functional recombinant human 4-1BB ligand for immune costimulatory therapy of cancer. J Immunother. 2011 Mar;34(2):175-82.

  [5]. Martinez-Perez AG, et al. 4-1BBL as a Mediator of Cross-Talk between Innate, Adaptive, and Regulatory Immunity against Cancer. Int J Mol Sci. 2021 Jun 9;22(12):6210.  [Content Brief]

  [6]. Salih HR, et al. Soluble CD137 (4-1BB) ligand is released following leukocyte activation and is found in sera of patients with hematological malignancies. J Immunol. 2001 Oct 1;167(7):4059-66.  [Content Brief]