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Mouse CD40 Ligand/TNFSF5 Antibody

Catalog #: AF1163
7 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF1163
AF1163-SP
CD40 Ligand/TNFSF5 in Transfected CHO-S (positive) and wild type (negative) cells.
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Citations (7)
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Mouse CD40 Ligand/TNFSF5 Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse CD40 Ligand/TNFSF5 in Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
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
Western Blot
0.1 µg/mL
Recombinant Mouse CD40 Ligand/TNFSF5 (Catalog # 1163-CL)
Immunocytochemistry
5-15 µg/mL
See below
Neutralization
Measured by its ability to neutralize CD40 Ligand/TNFSF5-induced proliferation in mouse splenic B cells. The Neutralization Dose (ND50) is typically 0.08-0.4 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

Immunocytochemistry CD40 Ligand/TNFSF5 antibody in transfected CHO-S cells by Immunocytochemistry (ICC). View Larger

CD40 Ligand/TNFSF5 in Transfected CHO-S (positive) and wild type (negative) cells. CD40 Ligand/TNFSF5 was detected in immersion fixed transfected CHO-S (positive) and wild type (negative) cells using Goat Anti-Mouse CD40 Ligand/TNFSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1163) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Neutralization Proliferation Induced by CD40 Ligand/TNFSF5 and Neutralization by Mouse CD40 Ligand/TNFSF5 Antibody. View Larger

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 Goat Anti-Mouse CD40 Ligand/TNFSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1163). The ND50 is typically 0.08-0.4 µg/mL.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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. Murine 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
  1. Armitage, R.J. et al. (1992) Nature 357:80.
  2. Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
  3. Fabienne, P. et al. (1996) J. Biol. Chem. 271:5965.
  4. Fabienne, P. et al. (1996) Eur. J. Immunol 26:725.
  5. Arrufo, A. et al. (1993) Cell 72:291.
  6. Hill, A. and N. Chapel et al. (1993) Nature 361:494.
  7. Korthauer, U. et al. (1993) Nature 361:539.
  8. Spriggs, M.K. et al. (1992) J. Exp. Med. 176:1543.
  9. Fanslow, W.C. et al. (1994) Seminars in Immunology 6:267.
  10. 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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Citations 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.

7 Citations: Showing 1 - 7
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  1. CD40 Enhances Type I Interferon Responses Downstream of CD47 Blockade, Bridging Innate and Adaptive Immunity
    Authors: S de Silva, G Fromm, CW Shuptrine, K Johannes, A Patel, KJ Yoo, K Huang, TH Schreiber
    Cancer Immunol Res, 2019-12-18;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Western Blot
  2. Identification and characterisation of the CD40-ligand of Sigmodon hispidus
    Authors: MS Russell, A Muralidhar, L Larocque, J Cao, Y Deschambau, J Varga, SN Thulasi Ra, X Li
    PLoS ONE, 2018-07-27;13(7):e0199067.
    Species: Mouse
    Sample Types: Recombinant Protein
    Applications: ELISA Development
  3. Vaccination with a fusion protein that introduces HIV-1 gag antigen into a multitrimer CD40L construct results in enhanced CD8+ T cell responses and protection from viral challenge by vaccinia-gag.
    Authors: Gupta, Sachin, Termini, James M, Raffa, Francesc, Williams, Cindi-An, Kornbluth, Richard, Stone, Geoffrey
    J Virol, 2013-11-13;88(3):1492-501.
    Species: Mouse
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  4. The tumor necrosis factor family receptors RANK and CD40 cooperatively establish the thymic medullary microenvironment and self-tolerance.
    Authors: Akiyama T, Shimo Y, Yanai H, Qin J, Ohshima D, Maruyama Y, Asaumi Y, Kitazawa J, Takayanagi H, Penninger JM, Matsumoto M, Nitta T, Takahama Y, Inoue J
    Immunity, 2008-09-19;29(3):423-37.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P, Neutralization
  5. CD40L expressed from the canarypox vector, ALVAC, can boost immunogenicity of HIV-1 canarypox vaccine in mice and enhance the in vitro expansion of viral specific CD8+ T cell memory responses from HIV-1-infected and HIV-1-uninfected individuals.
    Authors: Liu J, Yu Q, Stone GW, Yue FY, Ngai N, Jones RB, Kornbluth RS, Ostrowski MA
    Vaccine, 2008-06-02;26(32):4062-72.
    Species: Mouse
    Sample Types: Recombinant Protein
    Applications: Western Blot
  6. Preformed CD40 ligand exists in secretory lysosomes in effector and memory CD4+ T cells and is quickly expressed on the cell surface in an antigen-specific manner.
    Authors: Koguchi Y, Thauland TJ, Slifka MK, Parker DC
    Blood, 2007-06-26;110(7):2520-7.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  7. Reduced pathology following infection with transgenic Leishmania major expressing murine CD40 ligand.
    Authors: Field AE, Wagage S, Conrad SM
    Infect. Immun., 2007-04-02;75(6):3140-9.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: ELISA Development

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