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CD40L/CD154/TRAP Protein, Human (His)

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Based on 2 publication(s) in Google Scholar

CD40L (CD154; TRAP) is a ligand to CD40/TNFRSF5, acts function by generating a costimulatory signal that up-regulates IL-4 synthesis. CD40L is specifically expressed on activated CD4+ T-lymphocytes and involves in activation of NF-κB/MAPK pathway. CD40L also involves in B cell differentiation, maturation, and apoptosis. CD40L in human, cleaved into 2 chains of membrane form (1-261 a.a.) and soluble form (113-261 a.a.) which serves as a ligand for integrins (ITGA5:ITGB1 and ITGAV:ITGB3). CD40L/CD154/TRAP Protein, Human (His) has a total length of 144 amino acids (E108-L261), is expressed in E. coli cells with C-terminal His-tag.

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  • Species: Human
  • Source: E. coli
  • Storage:
    Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.
  • Experimental Validation Results for CD40L/CD154/TRAP Protein, Human (His)
  • CD40L/CD154/TRAP Protein, Human (His) Product Image
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  • Biological Activity
  • Technical Parameters
  • Product Properties
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  • References
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Biological Activity

Description

CD40L (CD154; TRAP) is a ligand to CD40/TNFRSF5, acts function by generating a costimulatory signal that up-regulates IL-4 synthesis[1]. CD40L is specifically expressed on activated CD4+ T-lymphocytes and involves in activation of NF-κB/MAPK pathway[2][3]. CD40L also involves in B cell differentiation, maturation, and apoptosis[4]. CD40L in human, cleaved into 2 chains of membrane form (1-261 a.a.) and soluble form (113-261 a.a.) which serves as a ligand for integrins (ITGA5:ITGB1 and ITGAV:ITGB3). CD40L/CD154/TRAP Protein, Human (His) has a total length of 144 amino acids (E108-L261), is expressed in E. coli cells with C-terminal His-tag.

Background

CD40 Ligand (CD40L; CD154; TRAP) belongs to the tumor necrosis factor (TNF) family, is the ligand for CD40/TNFRSF5, specifically expressed on activated CD4+ T-lymphocytes[1].
CD40L is a type II transmembrane protein on B cells triggers important signals for B cell differentiation, maturation, and apoptosis[4].
CD40L acts function by cross-linking on T-cells to generate a costimulatory signal and thus enhances the production of IL4 and IL10 in conjunction with the TCR/CD3 ligation and CD28 costimulation, as well as promoting the production of interferon-γ, and TNF-α[1][4].
CD40L, binding with CD40 on antigen-presenting cells (APC), activates TNFR-associated factor 2- and IKK2-dependent pathways with stimulating I-κB kinase (IKK), increasing NF-κB DNA binding, and p65 nuclear translocation. The activation of I-κB kinase leads to strongly c-Jun N-terminal kinase activation as well as GST-I-κB and GST-p65 phosphorylation[2].
CD40L involves in MAPK pathways that strongly repress Bcl-6 with inducing the phosphorylation of Erk1/2, p38 and Jnk1/2 and activating IRF4 mediated by NF-κB[3].
CD40L also binds to and signals through several integrins, including αvβ3 and α5β1, which bind to the trimeric interface of CD40L. CD40L plays a major role in immune response and is a major target for inflammation[5].
CD40L is widely found in different animals, while the sequence in Human is highly similar to Rhesus macaque (98.08%), but very different from Rat and Mouse with similarities of 77.31% and 77.69%, respectively. CD40L in Human is cleaved into 2 chains of membrane form (1-261 a.a.) and soluble form (113-261 a.a.), while the soluble form derives from the membrane form by proteolytic processing. Release of soluble CD40L from platelets is partially regulated by GP IIb/IIIa, actin polymerization, and a matrix metalloproteinases (MMP) inhibitor-sensitive pathway[6].

In Vitro

CD4 Ligand (2.86 ng/mL or 2.9 ng/mL) has been demonstrated to be a mark of cardiovascular risk, while the level (2.86 ng/mL) in who subsequently developed myocardial infarction, stroke, or cardiovascular death (cases) is higher than control group (2.9 ng/mL)[8].

In Vivo

CD4 Ligand (1 ng/mL; 24 hr) induces endothelial cell vascular cell adhesion molecule-1 (VCAM-1) expression. CD4 Ligand shows costimulation effect with IL-4 against VCAM-1[7].
CD4 Ligand (1 ng/mL; 24 hr) combined with IL-4 enhances IL-6 and inhibits IL-8 production by HUVEC[7].

Verified Bioactivity

1.Immobilized Human CD40 Ligand, His Tag at 5 μg/mL (100 μl/well) on the plate. Dose response curve for Human CD40, hFc Tag with the EC50 of ≤0.94 μg/mL determined by ELISA.
2. Immobilized Human CD40, hFc Tag at 2 μg/ml (100μl/well) on the plate. Dose response curve for Human/Cynomolgus/Rhesus macaque CD40 Ligand, His Tag with the EC50 of 57.6ng/ml determined by ELISA.

Technical Parameters

  • Species Human
  • Source E. coli
  • Tag C-8*His
  • Accession

    P29965 (Q114-L261)

  • Gene ID
    959  [NCBI]
  • Molecular Construction
    • N-term
    • CD40L (Q114-L261)
      Accession # P29965
    • 8*His
    • C-term
  • Protein Length

    Full Length of CD40L soluble form

  • Synonyms

    CD40LG; T-BAM; Prev. TNFSF5; Tumor Necrosis Factor (Ligand) Superfamily Member 5; Prev. HIGM1; Tumor Necrosis Factor Ligand Superfamily Member 5; Prev. IMD3; T-B Cell-Activating Molecule; CD40-L; CD40 Antigen Ligand; CD40L; T-Cell Antigen Gp39; TRAP; Tumo

  • AA Sequence

    QKGDQNPQIAAHVISEASSKTTSVLQWAEKGYYTMSNNLVTLENGKQLTVKRQGLYYIYAQVTFCSNREASSQAPFIASLCLKSPGRFERILLRAANTHSSAKPCGQQSIHLGGVFELQPGASVFVNVTDPSQVSHGTGFTSFGLLKL

  • Predicted Molecular Mass

    17.2 kDa

  • Molecular Weight

    Approximately 17 kDa, based on SDS-PAGE under reducing conditions.

  • Purity

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

Product Properties

Appearance

Solution

Formulation

Supplied as a 0.22 μm filtered solution of PBS, pH 7.4, 10% glycerol.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Storage & Stability

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping

Shipping with dry ice.

References

[1]. Blotta MH, et al. Cross-linking of the CD40 ligand on human CD4+ T lymphocytes generates a costimulatory signal that up-regulates IL-4 synthesis. J Immunol. 1996 May 1;156(9):3133-40. [Content Brief]

[2]. Schwabe RF, et al. CD40 activates NF-kappa B and c-Jun N-terminal kinase and enhances chemokine secretion on activated human hepatic stellate cells. J Immunol. 2001 Jun 1;166(11):6812-9. [Content Brief]

[3]. Batlle A, et al. CD40 and B-cell receptor signalling induce MAPK family members that can either induce or repress Bcl-6 expression. Mol Immunol. 2009 May;46(8-9):1727-35. [Content Brief]

[4]. Mikolajczak SA, et al. The modulation of CD40 ligand signaling by transmembrane CD28 splice variant in human T cells. J Exp Med. 2004 Apr 5;199(7):1025-31. [Content Brief]

[5]. Takada YK, et al. Soluble CD40L activates soluble and cell-surface integrin αvβ3, α5β1, and α4β1 by binding to the allosteric ligand-binding site (site 2). J Biol Chem. 2021 Jan-Jun;296:100399. [Content Brief]

[6]. Pietravalle F, et al. Human native soluble CD40L is a biologically active trimer, processed inside microsomes. J Biol Chem. 1996 Mar 15;271(11):5965-7. [Content Brief]

[7]. Kotowicz K, et al. Biological function of CD40 on human endothelial cells: costimulation with CD40 ligand and interleukin-4 selectively induces expression of vascular cell adhesion molecule-1 and P-selectin resulting in preferential adhesion of lymphocytes. Immunology. 2000 Aug;100(4):441-8. [Content Brief]

[8]. Schönbeck U, et al. Soluble CD40L and cardiovascular risk in women. Circulation. 2001 Nov 6;104(19):2266-8. [Content Brief]

[9]. Lee N, et al. Comparison of Flow-cytometric Antibody Secreting Cell Assay and Mabtech Immunoglobulin ELISpot Assay. Transplant Proc. 2017 Jun;49(5):963-966. [Content Brief]

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

  • CD40L/CD154/TRAP Protein, Human (His) Product Image
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  • Experimental Validation Results for CD40L/CD154/TRAP Protein, Human (His)
  • CD40L/CD154/TRAP Protein, Human (His) Product Image