Recombinant Rat CD40 Ligand/TNFSF5 (HA-tag) Protein, CF

  
  • Purity
    >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
  • Endotoxin Level
    <0.10 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured in a cell proliferation assay using mouse splenic B cells in the presence of IL-4. Banchereau, J. et al. (1991) Science 251:70. The ED50 for this effect is 0.15-0.9 ng/mL in the presence of 0.1 μg/mL of the cross-linking antibody, Mouse Anti-Hemagglutinin/HA Peptide Monoclonal Antibody (Catalog # MAB060).
  • Source
    Chinese Hamster Ovary cell line, CHO-derived
    HA
    (YPYDVPDYA)
    GCN4-IZ GGGSGGGSGGGS Rat CD40 Ligand
    (Met112-Leu260
    Accession # Q9Z2V2
    N-terminus C-terminus
  • Accession #
  • N-terminal Sequence
    Analysis
    Tyr
  • Predicted Molecular Mass
    22 kDa
  • SDS-PAGE
    22 - 30 kDa, reducing conditions
Carrier Free
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
8414-CL/CF
 
8414-CL
Formulation Lyophilized from a 0.2 μm filtered solution in sterile PBS and NaCl.
Formulation Lyophilized from a 0.2 μm filtered solution in PBS and NaCl with BSA as a carrier protein.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Reconstitution Reconstitute at 100 μg/mL in steril PBS containing at least 0.1% human or bovine serum albumin.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Data Images
Recombinant Rat CD40 Ligand (Catalog# 8414-CL/CF) induces cell proliferation by mouse splenic B cells in the presence of IL-4.  The ED50 for this effect is 0.15-0.9 ng/mL in the presence of 0.1 μg/mL of the cross-linking antibody, Mouse Anti-Hemagglutinin/HA Peptide Monoclonal Antibody (Catalog # MAB060).
1 μg/lane of Recombinant Rat CD40 Ligand/TNFSF5 was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing R bands at 26, 28 kDa and NR bands at 26, 28 kDa.
Background: CD40 Ligand/TNFSF5
CD40 Ligand, also known as TNFSF5, CD154, TRAP, or gp39, is a 33-39 kDa type II transmembrane glycoprotein member of the TNF superfamily (1, 2). Mature rat CD40 Ligand consists of a 22 amino acid (aa) cytoplasmic domain, a transmembrane segment, and an 217 aa extracellular region. The extracellular domain of rat CD40 Ligand shares 75% and 93% amino acid (aa) sequence identity with the human and mouse proteins, respectively. CD40 Ligand is expressed as a homotrimer on platelets and activated T cells and B cells. It is up-regulated following stimulation of basophils, eosinophils, fibroblasts, mast cells, monocytes, natural killer cells, vascular endothelial cells, and smooth muscle cells. CD40 Ligand binds and activates CD40, which is expressed on the surface of B cells, dendritic cells, macrophages, monocytes, platelets, endothelial cells, and epithelial cells (3). The 18 kDa soluble form (aa 112-260)  arises from proteolytic processing and retains the ability to bind and activate CD40 (4, 5). Monomeric, dimeric, and trimeric forms of soluble CD40 Ligand bind to oligomeric CD40 on cell membranes (2). CD40 ligation by CD40 Ligand promotes B cell activation and T cell-dependent humoral responses (6, 7). CD40 Ligand dysregulation on T cells and antigen presenting cells contributes to the immune deficiency associated with HIV infection and AIDS (8, 9). It is also implicated in the pathology of multiple cardiovascular diseases including atherosclerosis, atherothrombosis, and restenosis (10, 11).
  • References:
    1. Armitage, R.J. et al. (1992) Nature 357:80.
    2. Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
    3. van Kooten, C. and J. Banchereau (1997) Curr. Opin. Immunol. 9:330.
    4. Graf, D. et al. (1995) Eur. J. Immunol. 25:1749.
    5. Mazzei, G.J. et al. (1995) J. Biol. Chem. 270:7025.
    6. Rickert, R.C. et al. (2011) Immunol. Rev. 244:115.
    7. Elgueta, R. et al. (2009) Immunol. Rev. 229:152.
    8. Kornbluth, R.S. (2000) J. Leukoc. Biol. 68:373.
    9. Chougnet, C. (2003) J. Leukoc. Biol. 74:702.
    10. Pamukcu, B. et al. (2011) Ann. Med. 43:331.
    11. Hassan, G.S. et al. (2012) Immunobiology 217:521.
  • Entrez Gene IDs:
    959 (Human); 21947 (Mouse); 84349 (Rat)
  • Alternate Names:
    CD154 antigen; CD154; CD40 antigen ligand; CD40 Ligand; CD40-L; CD40LG; CD40LIGM; gp39; hCD40L; HIGM1; T-B cell-activating molecule; T-BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TNFSF5IMD3; TRAP; TRAPtumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome); tumor necrosis factor (ligand) superfamily member 5; Tumor necrosis factor ligand superfamily member 5
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