Human CD40 Ligand/TNFSF5 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB617

R&D Systems, part of Bio-Techne

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Blockade of Receptor-ligand Interaction, CyTOF-ready, Flow Cytometry, Immunohistochemistry, Western Blot

Cited:

Bioassay, Blocking, Immunohistochemistry, Neutralization

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 40804

View all CD40 Ligand/TNFSF5 Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CD40 Ligand/TNFSF5
Leu50-Leu261
Accession # P29965

Specificity

Detects human CD40 Ligand/TNFSF5 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) CD27 Ligand, rhCD30 Ligand, or recombinant mouse CD40 Ligand is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Endotoxin Level

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

Scientific Data Images for Human CD40 Ligand/TNFSF5 Antibody

CD40 Ligand/TNFSF5 antibody in Human Tonsil by Immunohistochemistry (IHC-P).

CD40 Ligand/TNFSF5 in Human Tonsil.

CD40 Ligand/TNFSF5 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human CD40 Ligand/TNFSF5 Monoclonal Antibody (Catalog # MAB617) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Applications for Human CD40 Ligand/TNFSF5 Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.03-0.1 µg/mL of this antibody will block 50% of the binding of 2 ng/mL of biotinylated Recombinant Human CD40 Ligand to immobilized Recombinant Human CD40/TNFRSF5 Fc Chimera (Catalog # 1493‑CD) coated at 2 µg/mL (100 µL/well). At 1 μg/mL, this antibody will block >90% of the binding.

CyTOF-ready

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

Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood mononuclear cells treated with PMA and Ca2+ ionomycin

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Western Blot

1 µg/mL
Sample: Recombinant Human CD40 Ligand/TNFSF5 aa 108-261 (Catalog # 6245-CL)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 3 using MAB617 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

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.

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), now renamed TNFSF5 but also known as CD154, TRAP and gp39, is a 34‑39 kDa type II transmembrane glycoprotein that belongs to the TNF superfamily (1‑3). Human CD40L is 261 amino acids (aa) in length and consists of a 22 aa cytoplasmic domain, a 24 aa transmembrane segment, and a 215 aa extracellular region that consists of multiple beta-strands and one N‑linked glycosylation site (4, 5). Although carbohydrates are present, they are not necessary for activity (6). As with other TNF superfamily members, CD40L will exist as a trimer, both as a membrane bound and soluble form (6‑8). The soluble form is 18 kDa in size and about 150 aa in length, and arises from intracellular proteolytic processing. As a trimer, the soluble form is bioactive (7‑9). Multiple mutations and alternate splice forms of CD40L exist, often associated with pathology and leading to truncated or nontrimerizable forms of CD40L (8). While CD40L is a ligand for CD40, the ligation of CD40L by CD40 initiates bidirectional signaling in both CD40 and CD40L expressing cells (10). The extracellular region of human CD40L is 99%, 88%, 86%, 82%, 75% and 75% aa identical to the extracellular regions of CD40L in rhesus monkey, bovine, porcine, canine, mouse and rat, respectively. CD40L binds to both CD40 and to integrin  alphaIIb beta3 (CD41) (3, 11). In the cell membrane, it also associates with a unique splice variant of CD28 (CD28i) that may facilitate CD40L signal transduction (12). CD40L is expressed by monocytes, NK cells, mast cells, basophils, smooth muscle cells, endothelial cells, dendritic cells, activated and resting B cells, plus activated platelets and CD4+ T cells (13, 14). CD40L ligation of CD40 on dendritic cells (DC) initiates DC maturation and differentiation. CD40L signaling into naïve B cells promotes germinal center formation and isotope switching. With IL-21, CD40L generates IgA plus IgG3; with IL-4, CD40L generates IgG1 secretion (14, 15).

References

  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) Prot. Exp. 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. Eissner, G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
  11. Prasad, K.S. et al. (2003) Proc. Natl. Acad. Sci. USA 100:12367.
  12. Mikolajczak, S.A. et al. (2004) J. Exp. Med. 199:1025.
  13. Lievens, D. et al. (2009) Thromb. Haemost. 102:206.
  14. Elgueta, R. et al. (2009) Immunol. Rev. 229:152.
  15. Avery, D.T. et al. (2008) J. Immunol. 181:1767.

Alternate Names

CD154, CD40L, CD40LG, gp39, TNFSF5, TRAP

Entrez Gene IDs

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

Gene Symbol

CD40LG

UniProt

P29965

Additional CD40 Ligand/TNFSF5 Products

Product Documents for Human CD40 Ligand/TNFSF5 Antibody

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Human CD40 Ligand/TNFSF5 Antibody

For research use only

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