Mouse CD40 Ligand/TNFSF5 Antibody

Catalog #: MAB1163
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB1163
MAB1163-SP

Detection of CD40 Ligand/TNFSF5 in Mouse Splenocytes by Flow Cytometry.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Mouse CD40 Ligand/TNFSF5 Antibody Summary

Species Reactivity

Mouse

Specificity

Detects mouse CD40 Ligand in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant mouse (rm) CD27 Ligand, rmCD30 Ligand, or recombinant human CD40 Ligand is observed.

Source

Monoclonal Rat IgG_2A Clone # 208109

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse CD40 Ligand/TNFSF5
Glu61-Leu260
Accession # P27548

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Endotoxin Level

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

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Neutralization

Measured by its ability to neutralize CD40 Ligand/TNFSF5-induced proliferation in mouse splenic B cells. The Neutralization Dose (ND₅₀) is typically 0.0075-0.045 µg/mL in the presence of 0.5 ng/mL Recombinant Mouse CD40 Ligand/TNFSF5.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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Detection of CD40 Ligand/TNFSF5 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes either (A) stimulated to induce Th1 cells or (B) unstimulated were stained with Rat Anti-Mouse CD40 Ligand/TNFSF5 Monoclonal Antibody (Catalog # MAB1163) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113) and Rat Anti-Mouse CD4 PE-conjugated Monoclonal Antibody (Catalog # FAB554P). Quadrant markers were set based on control antibody staining (Catalog # MAB006). View our protocol for Staining Membrane-associated Proteins.

Neutralization

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Proliferation Induced by CD40 Ligand/TNFSF5 and Neutralization by Mouse CD40 Ligand/TNFSF5 Antibody. Recombinant Mouse CD40 Ligand/TNFSF5 (Catalog # 8230-CL) induces proliferation in mouse splenic B cells in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse CD40 Ligand/TNFSF5 (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse CD40 Ligand/TNFSF5 Monoclonal Antibody (Catalog # MAB1163). The ND₅₀ is typically 0.0075-0.045 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CD40 Ligand/TNFSF5

CD40 ligand (CD40L; also known as CD154, TNFSF5, TRAP, or gp39) is a 260 amino acid (aa) type II transmembrane glycoprotein belonging to the TNF family. Mouse CD40L consists of a 22 aa cytoplasmic domain, a 24 aa transmembrane domain, and 214 aa extracellular domain bearing a single glycosylation site (1, 2). CD40L is expressed predominantly on activated CD4⁺ T lymphocytes and also found in other types of cells, including NK cells, mast cells, basophils, and eosinophils. Murine CD40L shares 78% amino acid sequence identity with human CD40L. Native bioactive soluble CD40L exists. Soluble human trimeric CD40L secreted by stimulated T cells has been shown to be generated by proteolysis in the microsomes (3). Both membrane bound and soluble CD40L induce similar effects on B cells (3, 4). The receptor of CD40L is CD40, a type I transmembrane glycoprotein belonging to the TNF receptor family. CD40 is expressed on B lymphocytes, monocytes, dendritic cells, and thymic epithelium. Although all monomeric, dimeric, and trimeric forms of soluble CD40L can bind to CD40, the soluble trimeric form of CD40L has the most potent biological activity through oligomerization of cell surface CD40, a common feature of TNF receptor family members (2). The genetic defect in the hyper-IgM syndrome is due to point mutations or deletions of the gene encoding the CD40L, which prevent CD40L from interacting with CD40 (5‑7). CD40L mediates a range of activities on B cells including induction of activation-associated surface antigen, entry into the cell cycle, isotype switching, Ig secretion, and memory generation (8, 9). CD40-CD40L interaction also plays important roles in monocyte activation and dendritic cell maturation (10).

References

Armitage, R.J. et al. (1992) Nature 357:80.
Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
Fabienne, P. et al. (1996) J. Biol. Chem. 271:5965.
Fabienne, P. et al. (1996) Eur. J. Immunol 26:725.
Arrufo, A. et al. (1993) Cell 72:291.
Hill, A. and N. Chapel et al. (1993) Nature 361:494.
Korthauer, U. et al. (1993) Nature 361:539.
Spriggs, M.K. et al. (1992) J. Exp. Med. 176:1543.
Fanslow, W.C. et al. (1994) Seminars in Immunology 6:267.
Kooten, C.V. and J. Banchereau (2000) J. Leukoc. Biol. 67:2.

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

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Citation for Mouse CD40 Ligand/TNFSF5 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

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