Recombinant Human CD40 Ligand/TNFSF5 (108-261), Animal-Free

In a functional ELISA, Animal-Free™ Recombinant Human CD40 Ligand/TNFSF5 (aa 108-261) (Catalog # AFL617) binds to Recombinant Human CD40/TNFRSF5 Fc Chimera (1493-CDB) with an ED50 of 0.300-3.60 ng/mL.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human CD40 Ligand/TNFSF5 (108-261), Animal-Free Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. Animal-Free™ Recombinant Human CD40 Ligand/TNFSF5 aa 108-261 (Catalog # AFL617) binds to Recombinant Human CD40/TNFRSF5 Fc Chimera (Catalog # 1493-CDB) with an ED 50 of 0.300-3.60 ng/mL.
• Source: E. coli-derived human CD40 Ligand/TNFSF5 protein Glu108-Leu261, with an N-terminal Met Produced using non-animal reagents in an animal-free laboratory.
• Accession #: P29965
• N-terminal Sequence: Met
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• Gene: CD40LG
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 16.9 kDa (monomer).
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 18 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 3 months, 2 to 8 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in NaH 2 PO 4 NaCl and EDTA.
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human CD40 Ligand/TNFSF5 (108-261), Animal-Free

• CD154 antigen
• CD154
• CD40 antigen ligand
• CD40 Ligand
• CD40L
• 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

Background

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).

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.

Manufacturing Process

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

• All molecular biology procedures use animal-free media and dedicated labware.
• Dedicated fermentors are utilized in committed animal-free areas.

Purification

• Protein purification columns are animal-free.
• Bulk proteins are filtered using animal-free filters.
• Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

• Low Endotoxin Level.
• No impairment of biological activity.
• High quality product obtained under stringent conditions.
• For ex vivo research or bioproduction, additional documentation can be provided.

Please read our complete Animal-Free Statement

 

FAQs for CD40 Ligand/TNFSF5 (AFL617). (Showing 1 - 1 of 1 FAQ).

1. I am looking for paired ABs for use in detecting CD40L (rat) by ELISA. Do you have such a reagent(s) available?
   • Unfortunately we currently do not have any CD40 Ligand antibody pairs that have been validated in Rat. The only antibody pairs we offer for CD40L are validated in Human. I apologize for this inconvenience. If you would be interested in testing one of our products in Rat I'd like to invite you to participate in our Innovators Reward Program. Please contact us at innovators@novusbio.com with any questions regarding this program.

Bioinformatics

• Gene Symbol: CD40LG
• Uniprot:
  • P29965()