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

Catalog #: AFL617 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AFL617

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

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

Learn more about Animal-Free Recombinant Proteins

Animal Free Proteins
Animal-free proteins are particularly important for researchers concerned with experimental variables caused by trace animal components or mammalian pathogens. Our products generated under animal-free conditions share the same biological activities as those produced using our standard laboratory techniques.

Product Specifications

Purity

>97%, 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 B cell-enriched peripheral blood mononuclear cells (PBMC) in the presence of IL-4. Spriggs, M.K. et al. (1992) J. Exp. Med. 176:1543. The ED₅₀ for this effect is 1‑3 µg/mL in the presence of 20 ng/mL of recombinant human IL·4.

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
Analysis

Met

Predicted Molecular Mass

16.9 kDa (monomer)

SDS-PAGE

18 kDa, reducing conditions

Product Datasheets

AFL617

Product Datasheet

COA

SDS

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.

AFL617

Formulation	Lyophilized from a 0.2 μm filtered solution in NaH₂PO₄, NaCl and EDTA.
Reconstitution	Reconstitute at 0.2 mg/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. 3 months, 2 to 8 °C under sterile conditions after reconstitution.

Reconstitution Calculator

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

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

Manufacturing Specifications

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