Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged)

R&D Systems | Catalog # 2706-CL

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) (2706-CL)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P29965

Applications

Bioactivity
View all CD40 Ligand/TNFSF5 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human CD40 Ligand/TNFSF5 protein
Glu108-Leu261, with an N-terminal Met and 6-His tag

Purity

>97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.01 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

17.7 kDa (monomer)

SDS-PAGE

18 kDa, reducing conditions

Activity

Measured in a cell proliferation assay using enriched human B cells in the presence of IL-4. Spriggs, M.K. et al. (1992) J. Exp. Med. 176:1543.
The ED50 for this effect is 2‑12 ng/mL in the presence of 10 µg/mL of a cross-linking antibody, Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050) and 20 ng/mL of recombinant human IL-4.

Reviewed Applications

Read 1 review rated 4 using 2706-CL in the following applications:

Scientific Data Images for Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged)

Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) Bioactivity

Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) Bioactivity

Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) (Catalog # 2706-CL) stimulates cell proliferation using enriched B cells in the presence of IL-4. The ED50 for this effect is 2-12 ng/mL in the presence of 10 µg/mL of a cross-linking antibody, Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050) and 20 ng/mL of Recombinant Human IL-4 (Catalog # 204-IL).

Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) SDS-PAGE

Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) SDS-PAGE

1 μg/lane of Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a single band at 19 kDa.

Formulation, Preparation, and Storage

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.

Carrier: 2706-CL
Formulation Lyophilized from a 0.2 μm filtered solution in HEPES, NaCl and EDTA with BSA as a carrier protein.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
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.
Carrier Free: 2706-CL/CF
Formulation Lyophilized from a 0.2 μm filtered solution in HEPES, NaCl and EDTA.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS.
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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CD40 Ligand/TNFSF5

CD40 Ligand, also known as TNFSF, CD154, TRAP, and gp39, is a 34-39 kDa type II transmembrane glycoprotein that belongs to the TNF superfamily (1-3). Mature human CD40 Ligand consists of a 22 amino acid (aa) cytoplasmic domain, a transmembrane segment, and an 215 aa extracellular region (4, 5). The extracellular domain of human CD40 Ligand shares 74% and 76% aa sequence identity with mouse and rat CD40 Ligand, respectively. Similar to other TNF superfamily members, CD40 Ligand forms a bioactive homotrimer, both as membrane bound and soluble forms (6-9). The 18 kDa soluble form (aa 113-261) arises from proteolytic processing. Mutation and alternative splicing generate additional forms of CD40 Ligand that are often truncated or non-trimerizable (8). CD40 Ligand is expressed on platelets, as well as on activated T cells and B cells, basophils, eosinophils, fibroblasts, mast cells, monocytes, natural killer cells, vascular endothelial cells, and smooth muscle cells. CD40 Ligand binds to CD40, which is expressed on the surface of B cells, dendritic cells, macrophages, monocytes, platelets, endothelial, and epithelial cells (10). The interaction of CD40 Ligand with CD40 initiates signaling in both CD40 and CD40 Ligand expressing cells (11).  CD40 ligation by CD40 Ligand promotes B cell activation and T cell-dependent humoral responses (12, 13). CD40 Ligand dysregulation on T cells and antigen presenting cells contributes to the immune deficiency associated with HIV infection and AIDS (14, 15). It is also implicated in the pathology of multiple cardiovascular diseases including atherosclerosis, atherothrombosis, and restenosis (16, 17).

References

  1. Zhang, G. (2004) Curr. Opin. Struct. Biol. 14:154.
  2. Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
  3. Quezada, S.A. et al. (2004) Annu. Rev. Immunol. 22:307.
  4. Graf, D. et al. (1992) Eur. J. Immunol. 22:3191.
  5. Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
  6. Khandekar, S.S. et al. (2001) Protein Expr. Purif. 23:301.
  7. Pietravalle, F. et al. (1996) J. Biol. Chem. 271:5965.
  8. Garber, E. et al. (1999) J. Biol. Chem. 274:33545.
  9. Vakkalanka, R.K. et al. (1999) Arthritis Rheum. 42:871.
  10. van Kooten, C. and J. Banchereau (1997) Curr. Opin. Immunol. 9:330.
  11. Eissner, G. et al. (2004) Cytokine Growth Factor. Rev. 15:353.
  12. Rickert, R.C. et al. (2011) Immunol. Rev. 244:115.
  13. Elgueta, R. et al. (2009) Immunol. Rev. 229:152.
  14. Kornbluth, R.S. (2000) J. Leukoc. Biol. 68:373.
  15. Chougnet, C. (2003) J. Leukoc. Biol. 74:702.
  16. Pamukcu, B. et al. (2011) Ann. Med. 43:331.
  17. Hassan, G.S. et al. (2012) Immunobiology 217:521.

Alternate Names

CD154, CD40L, CD40LG, gp39, TNFSF5, TRAP

Entrez Gene IDs

959 (Human); 21947 (Mouse); 84349 (Rat)

Gene Symbol

CD40LG

UniProt

P29965

Additional CD40 Ligand/TNFSF5 Products

Product Documents for Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS 2706-CL (with carrier)
Download SDS 2706-CL/CF (carrier free)

Product Specific Notices for Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged)

For research use only

Citations for Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged)

Customer Reviews for Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) (1)

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FAQs for Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged)

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    • Q: Is Recombinant Human CD40 Ligand/TNFSF5 (Histidine-tagged) (Catalog # 2706-CL) a homotrimer?
      A: While native CD40L is most active as a trimer, this recombinant protein may not fully achieve that configuration. The cross-linking ability of the anti-polyhistidine antibody used in the QC bioassay assists in this process to promote optimal activity. We do not have a structural specification for this product, relying instead on bioassay with cross-linking to confirm consistency in bioactivity.

    • Q: This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?
      A: The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.
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