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

R&D Systems, part of Bio-Techne | Catalog # 8414-CL

Analyzed by SEC-MALS
R&D Systems, part of Bio-Techne
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Carrier Free
8414-CL-025/CF
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With Carrier
8414-CL-025
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Key Product Details

Source

CHO

Accession #

Q9Z2V2

Conjugate

Unconjugated

Applications

Bioactivity

View all CD40 Ligand/TNFSF5 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived rat CD40 Ligand/TNFSF5 protein
HA
(YPYDVPDYA)		 GCN4-IZ GGGSGGGSGGGS Rat CD40 Ligand
(Met112-Leu260
Accession # Q9Z2V2
N-terminus C-terminus

Purity

>90%, by SDS-PAGE with silver staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Tyr

Predicted Molecular Mass

22 kDa

SDS-PAGE

22-30 kDa, reducing conditions

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).

Scientific Data Images for Recombinant Rat CD40 Ligand/TNFSF5 (HA-tag) Protein

Recombinant Rat CD40 Ligand/TNFSF5 Protein SEC-MALS.

Recombinant Rat CD40 Ligand/TNFSF5 (Catalog # 8414-CL) has a molecular weight (MW) of 67.1 kDa as analyzed by SEC-MALS, suggesting that this protein is a homotrimer.  MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
Recombinant Rat CD40 Ligand/TNFSF5 (HA-tag) Protein Bioactivity

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

Recombinant Rat CD40 Ligand (Catalog# 8414-CL) 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).
Recombinant Rat CD40 Ligand/TNFSF5 (HA-tag) Protein SDS-PAGE

Recombinant Rat CD40 Ligand/TNFSF5 (HA-tag) Protein SDS-PAGE

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.

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: 8414-CL
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 steril 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: 8414-CL/CF
Formulation Lyophilized from a 0.2 μm filtered solution in sterile PBS and NaCl.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Shipping The product is shipped with polar packs. 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.

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.

Alternate Names

CD154, CD40L, CD40LG, gp39, TNFSF5, TRAP

Entrez Gene IDs

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

Gene Symbol

CD40LG

UniProt

Q9Z2V2

Additional CD40 Ligand/TNFSF5 Products

Product Documents for Recombinant Rat CD40 Ligand/TNFSF5 (HA-tag) Protein

Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS 8414-CL (with carrier)
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Product Specific Notices for Recombinant Rat CD40 Ligand/TNFSF5 (HA-tag) Protein

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