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4-1BBL/TNFSF9 Protein, Human

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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). 4-1BBL stimulates T cell proliferation and induces effective anti-tumor immune responses. 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. 4-1BBL/TNFSF9 Protein, Human is a recombinant protein consisting of 184 amino acids (R71-E254) and is produced in E. coli.

For research use only. We do not sell to patients.
  • Species: Human
  • Source: E. coli
  • Storage:
    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.
  • Experimental Validation Results for 4-1BBL/TNFSF9 Protein, Human
  • 4-1BBL/TNFSF9 Protein, Human Product Image
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  • Biological Activity
  • Technical Parameters
  • Product Properties
  • Documentation
  • References
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Biological Activity

Description

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, Human is a recombinant protein consisting of 184 amino acids (R71-E254) and is produced in E. coli.

Background

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].

In Vitro

4-1BB (human; 1 μg/mL; 16 h) leads to induction of monocyte activation and induces Myc protein production[4].
4-1BB (human; 1 μg/mL; 1 d) prolongs survival of peripheral monocytes and (1 μg/mL; 1 h) induces expression of M-CSF[5].
4-1BB (human; 1.2-4 μg/mL; 7 d) exhibits nhancement of cell numbers dose-dependently[6].

Verified Bioactivity

Immobilized Recombinant Cynomolgus 4-1BB (C-Fc) at 1 μg/mL (100 μL/well) can bind Recombinant Human 4-1BBL (C-6His). The ED50 of Recombinant Human 4-1BBL (C-6His) is 30-60 ng/mL. 

Technical Parameters

  • Species Human
  • Source E. coli
  • Tag C-6*His
  • Accession

    P41273 (R71-E254)

  • Gene ID
    8744  [NCBI]
  • Molecular Construction
    • N-term
    • 4-1BBL (R71-E254)
      Accession # P41273
    • 6*His
    • C-term
  • Protein Length

    Partial

  • Synonyms

    TNFSF9; Homolog Of Mouse 4-1BB-L; TNF Superfamily Member 9; Receptor 4-1BB Ligand; 4-1BBL; CD137 Ligand; 4-1BB-L; 4-1BB Ligand; CD137L; Tumor Necrosis Factor (Ligand) Superfamily, Member 9, Isoform CRA_a; Tumor Necrosis Factor (Ligand) Superfamily, Member

  • AA Sequence

    REGPELSPDDPAGLLDLRQGMFAQLVAQNVLLIDGPLSWYSDPGLAGVSLTGGLSYKEDTKELVVAKAGVYYVFFQLELRRVVAGEGSGSVSLALHLQPLRSAAGAAALALTVDLPPASSEARNSAFGFQGRLLHLSAGQRLGVHLHTEARARHAWQLTQGATVLGLFRVTPEIPAGLPSPRSE

  • Predicted Molecular Mass

    20.6 kDa

  • Molecular Weight

    Approximately 16-20 kDa, based on SDS-PAGE under reducing conditions.

  • Purity

    ≥ 95%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Lyophilized powder.

Formulation

Lyophilized from a 0.22 μm filtered solution of 20 mM Tris-HCl, 150 mM NaCl, pH 7.4.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

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).

Storage & Stability

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.

Shipping

Room temperature in continental US; may vary elsewhere.

References

[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]. Tan JT, et al. 4-1BB ligand, a member of the TNF family, is important for the generation of antiviral CD8 T cell responses. J Immunol. 1999 Nov 1;163(9):4859-68. [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]. Cannons JL, et al. 4-1BB ligand induces cell division, sustains survival, and enhances effector function of CD4 and CD8 T cells with similar efficacy. J Immunol. 2001 Aug 1;167(3):1313-24. [Content Brief]

[5]. 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.

[6]. Cheuk AT, et al. Role of 4-1BB:4-1BB ligand in cancer immunotherapy. Cancer Gene Ther. 2004 Mar;11(3):215-26. [Content Brief]

[7]. 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]

[8]. 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]

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Calculators

Reconstitution Calculator

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Dilution Calculator

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The Specific Activity Calculator Equation
  • Specific Activity (Unit/mg)
  • Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) Specific Activity (Unit/mg)
Unit/mg
= 106 ÷
Biological Activity (ED50) Biological Activity (ED50)
106 ÷
ng/mL
MOQ
Minimum order quantity
100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     ≥ 95%, as determined by reducing SDS-PAGE.

  • 4-1BBL/TNFSF9 Protein, Human Product Image
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  • Experimental Validation Results for 4-1BBL/TNFSF9 Protein, Human
  • 4-1BBL/TNFSF9 Protein, Human Product Image